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<a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science/journal/01429612"><b>Biomaterials</b></a><br>
<a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=PublicationURL&_tockey=%23TOC%235558%232005%23999739998%23505012%23FLA%23&_cdi=5558&_pubType=J&view=c&_auth=y&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=e889bb525e72c7ce462ca69c1fd48125">
Volume 26, Issue 1</a>,
January 2005,
Pages 93-99
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return false;" onclick="var moreDiv=document.getElementById('nbApitagCloudtreatment'); return nbApi.onEvent(event,{owner:this,redir:true,type:'treatment',viewMore: moreDiv!=null?moreDiv.nbExpand:'no',sessionId:nbApi.sessionId,pii:'37175',strategy:'CLOUD_BY_TYPE',size:5,containerId:'nextBioViewer',tagName:'l-glutamate',customParam:'logId=' + nbApi.sessionId + '&tcid=88c196b8deb74a77b3eafb2388aa7c02&log=true&logTagType=treatment&logTagId=37175&logTagName=l-glutamate&it=Inkjet%20printing%20of%20viable%20mammalian%20cells&dpId=1&type=treatment&id=37175&q=L-Glutamic Acid&bbut=false'});">l-glutamate</a> </td> <td> <a href="" target="_blank" onmouseover="nbApi.onEvent(event, 'nextBioViewer', 'l-glutamine'); return false;" onclick="var moreDiv=document.getElementById('nbApitagCloudtreatment'); return nbApi.onEvent(event,{owner:this,redir:true,type:'treatment',viewMore: moreDiv!=null?moreDiv.nbExpand:'no',sessionId:nbApi.sessionId,pii:'36607',strategy:'CLOUD_BY_TYPE',size:5,containerId:'nextBioViewer',tagName:'l-glutamine',customParam:'logId=' + nbApi.sessionId + '&tcid=88c196b8deb74a77b3eafb2388aa7c02&log=true&logTagType=treatment&logTagId=36607&logTagName=l-glutamine&it=Inkjet%20printing%20of%20viable%20mammalian%20cells&dpId=1&type=treatment&id=36607&q=Glutamine&bbut=false'});">l-glutamine</a> </td> </tr> <tr class="nbApiTagCloudMore"> <td> <a href="" target="_blank" onmouseover="nbApi.onEvent(event, 'nextBioViewer', 'sodium bicarbonate'); return false;" onclick="var moreDiv=document.getElementById('nbApitagCloudtreatment'); return nbApi.onEvent(event,{owner:this,redir:true,type:'treatment',viewMore: moreDiv!=null?moreDiv.nbExpand:'no',sessionId:nbApi.sessionId,pii:'25288',strategy:'CLOUD_BY_TYPE',size:5,containerId:'nextBioViewer',tagName:'sodium bicarbonate',customParam:'logId=' + nbApi.sessionId + '&tcid=88c196b8deb74a77b3eafb2388aa7c02&log=true&logTagType=treatment&logTagId=25288&logTagName=sodium bicarbonate&it=Inkjet%20printing%20of%20viable%20mammalian%20cells&dpId=1&type=treatment&id=25288&q=Sodium bicarbonate&bbut=false'});">sodium bicarbonate</a> </td> </tr></tbody></table> <div class="nbApiTagCloudMoreLink" style="clear: both;"> <img src="science-direct_files/i_Next.gif" alt=""> <a class="nbApiTagCloudLess" onclick='nbApi.tagCloudShowMore("treatment")'>View more...</a> <a class="nbApiTagCloudMore" onclick='nbApi.tagCloudShowLess("treatment")'>View less...</a> </div> <div style="clear: both;"></div> </div> <div class="nbApiTagCloudGroup nbApiTagCloudShowLess" id="nbApitagCloudtissue"> <div class="nbApiTagCloudGroupHead"> <img src="science-direct_files/tissue.png" alt=""> tissues<span class="nbApiTagCloudGroupCount"> | 10</span> </div> <table cellpadding="0" cellspacing="0"> <tbody><tr> <td width="50%"> <a href="" target="_blank" onmouseover="nbApi.onEvent(event, 'nextBioViewer', 'motoneurons'); 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return false;" onclick="var moreDiv=document.getElementById('nbApitagCloudtissue'); return nbApi.onEvent(event,{owner:this,redir:true,type:'tissue',viewMore: moreDiv!=null?moreDiv.nbExpand:'no',sessionId:nbApi.sessionId,pii:'128789',strategy:'CLOUD_BY_TYPE',size:10,containerId:'nextBioViewer',tagName:'stem cells',customParam:'logId=' + nbApi.sessionId + '&tcid=88c196b8deb74a77b3eafb2388aa7c02&log=true&logTagType=tissue&logTagId=128789&logTagName=stem cells&it=Inkjet%20printing%20of%20viable%20mammalian%20cells&dpId=1&type=tissue&id=128789&q=Stem cell&bbut=false'});">stem cells</a> </td> <td> <a href="" target="_blank" onmouseover="nbApi.onEvent(event, 'nextBioViewer', 'stomachs'); return false;" onclick="var moreDiv=document.getElementById('nbApitagCloudtissue'); return nbApi.onEvent(event,{owner:this,redir:true,type:'tissue',viewMore: moreDiv!=null?moreDiv.nbExpand:'no',sessionId:nbApi.sessionId,pii:'103989',strategy:'CLOUD_BY_TYPE',size:10,containerId:'nextBioViewer',tagName:'stomachs',customParam:'logId=' + nbApi.sessionId + '&tcid=88c196b8deb74a77b3eafb2388aa7c02&log=true&logTagType=tissue&logTagId=103989&logTagName=stomachs&it=Inkjet%20printing%20of%20viable%20mammalian%20cells&dpId=1&type=tissue&id=103989&q=Stomach&bbut=false'});">stomachs</a> </td> </tr> </tbody></table> <div class="nbApiTagCloudMoreLink" style="clear: both;"> <img src="science-direct_files/i_Next.gif" alt=""> <a class="nbApiTagCloudLess" onclick='nbApi.tagCloudShowMore("tissue")'>View more...</a> <a class="nbApiTagCloudMore" onclick='nbApi.tagCloudShowLess("tissue")'>View less...</a> </div> <div style="clear: both;"></div> </div> <div class="nbApiTagCloudGroup nbApiTagCloudShowLess nbApiTagCloudGroupLast" id="nbApitagCloudbiogroup"> <div class="nbApiTagCloudGroupHead"> <img src="science-direct_files/biogroup.png" alt=""> biogroups<span class="nbApiTagCloudGroupCount"> | 1</span> </div> <table cellpadding="0" cellspacing="0"> <tbody><tr> <td width="50%"> <a href="" target="_blank" onmouseover="nbApi.onEvent(event, 'nextBioViewer', 'lactate dehydrogenase'); return false;" onclick="var moreDiv=document.getElementById('nbApitagCloudbiogroup'); return nbApi.onEvent(event,{owner:this,redir:true,type:'biogroup',viewMore: moreDiv!=null?moreDiv.nbExpand:'no',sessionId:nbApi.sessionId,pii:'200455',strategy:'CLOUD_BY_TYPE',size:1,containerId:'nextBioViewer',tagName:'lactate dehydrogenase',customParam:'logId=' + nbApi.sessionId + '&tcid=88c196b8deb74a77b3eafb2388aa7c02&log=true&logTagType=biogroup&logTagId=200455&logTagName=lactate dehydrogenase&it=Inkjet%20printing%20of%20viable%20mammalian%20cells&dpId=1&type=biogroup&id=200455&q=Lactate dehydrogenase&bbut=false'});"><span title="lactate dehydrogenase">lactate dehydrogenas…</span></a> </td> </tr></tbody></table> </div> </div></div> <div class="nbApiBottom"><div class="nbApiFoot">Scientific correlations by NextBio</div></div> </div></div></div>
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<tbody><tr><td scope="col" class="greyHeader" style="background-image: url(/scidirimg/right_gradient.gif);"><b>Related Articles in ScienceDirect</b></td></tr></tbody><tbody id="mlkt"><tr><td><div id="mlktList"><div class="infobubble-container"><div class="mlktLink" id="mlktLink_1"><table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td style="padding: 5px 5px 0px;" valign="top"><img src="science-direct_files/bullet_square.gif" alt=""></td><td onmouseover="InfoBubble.show('infobubble_1','mlktLink_1','iframe_1')" onmouseout="InfoBubble.timeout()"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6WN5-4GSCHN7-8&_user=108429&_origUdi=B6TWB-4CGM8MB-1&_fmt=high&_coverDate=04%2F30%2F1991&_rdoc=1&_orig=article&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=fdc7f877ddeaa33f6bf260332bdc7efe">Expression of antibody genes in nonlymphoid mammalian c...<br><i>Methods</i></a><br><iframe id="iframe_1" style="display: none; position: absolute; z-index: 1;" scrolling="no" frameborder="0"></iframe><div class="infobubble" id="infobubble_1"><table cellpadding="0" cellspacing="0" width="100%"><tbody><tr><td><div style="float: right; padding-left: 5px;"><a onclick="InfoBubble.hide()"><img src="science-direct_files/btn_xclose.gif" alt="Close" title="Close" onmouseover="javascript:this.src='/scidirimg/btn_xclose_hov.gif';" onmouseout="javascript:this.src='/scidirimg/btn_xclose.gif';"></a></div><img src="science-direct_files/jrn_sub.gif" alt="You are entitled to access the full text of this document" title="You are entitled to access the full text of this document" border="0" width="12" height="14"> <b><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6WN5-4GSCHN7-8&_user=108429&_coverDate=04%2F30%2F1991&_rdoc=1&_fmt=high&_orig=article&_cdi=6953&_sort=v&_docanchor=&view=c&_ct=1512&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=e2d82ab98abae7f909a5532814286a3e">Expression of antibody genes in nonlymphoid mammalian cells</a></b><br><i>Methods</i>, <i>Volume 2, Issue 2</i>, <i>April 1991</i>, <i>Pages 136-145</i><br>Christopher R. Bebbington<br><br><b>Abstract</b><br><div class="mlktScroll"><div style="line-height: 150%;">A
wide variety of mammalian cell lines have the ability to assemble and
secrete immunoglobulins. This provides the possibility of adapting
highly efficient mammalian cell expression systems available for
nonlymphoid host cells to obtain efficient expression of recombinant
antibodies. A set of vectors that use a powerful cytomegalovirus
promoter enhancer to direct expression of the coding sequence for each
immunoglobulin chain is described. The vectors also contain a
replication origin from SV40 to generate high copy numbers in COS
monkey kidney cells and a marker gene for selection of permanently
transfected cell lines. Methods for generating microgram amounts of
antibody in 2–4 days from COS cells and for stable introduction of the
vectors into CHO cells are described. The use of a glutamine synthetase
(GS) selectable marker to obtain vector amplification in CHO cells is
also described. This permits the generation of cell lines that can
accumulate in excess of 200 mg/liter of antibody in the culture medium.</div></div><br><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MImg&_imagekey=B6WN5-4GSCHN7-8-1&_cdi=6953&_user=108429&_orig=article&_coverDate=04%2F30%2F1991&_sk=999979997&view=c&wchp=dGLbVzW-zSkWA&md5=21ef2557f75535cd0ac3620658c52143&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (1032 K)</a>
</td></tr></tbody></table></div></td></tr></tbody></table></div><div class="infobubble-container"><div class="mlktLink" id="mlktLink_2"><table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td style="padding: 5px 5px 0px;" valign="top"><img src="science-direct_files/bullet_square.gif" alt=""></td><td onmouseover="InfoBubble.show('infobubble_2','mlktLink_2','iframe_2')" onmouseout="InfoBubble.timeout()"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B73F6-47S6466-3F&_user=108429&_origUdi=B6TWB-4CGM8MB-1&_fmt=high&_coverDate=10%2F15%2F1995&_rdoc=1&_orig=article&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=ca4543a6eb0d4833f3c1534fff96aa87">Expression of endogenous muscarinic acetylcholine recep...<br><i>European Journal of Pharmacology: Molecular Pharmacolog...</i></a><br><iframe id="iframe_2" style="display: none; position: absolute; z-index: 1;" scrolling="no" frameborder="0"></iframe><div class="infobubble" id="infobubble_2"><table cellpadding="0" cellspacing="0" width="100%"><tbody><tr><td><div style="float: right; padding-left: 5px;"><a onclick="InfoBubble.hide()"><img src="science-direct_files/btn_xclose.gif" alt="Close" title="Close" onmouseover="javascript:this.src='/scidirimg/btn_xclose_hov.gif';" onmouseout="javascript:this.src='/scidirimg/btn_xclose.gif';"></a></div><img src="science-direct_files/jrn_sub.gif" alt="You are entitled to access the full text of this document" title="You are entitled to access the full text of this document" border="0" width="12" height="14"> <b><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B73F6-47S6466-3F&_user=108429&_coverDate=10%2F15%2F1995&_rdoc=1&_fmt=high&_orig=article&_cdi=11490&_sort=v&_docanchor=&view=c&_ct=1512&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=db25c9b74839daf72d79d9f56d82ecd7">Expression of endogenous muscarinic acetylcholine receptors in Chinese hamster ovary cells</a></b><br><i>European Journal of Pharmacology: Molecular Pharmacology</i>, <i>Volume 291, Issue 2</i>, <i>15 October 1995</i>, <i>Pages R1-R2</i><br>Shou Zhen Wang, Sheng Zu Zhu, Esam E. El-Fakahany<br><br><b>Abstract</b><br><div class="mlktScroll"><div style="line-height: 150%;">Chinese
hamster ovary (CHO) cells are commonly used for expression of the genes
of cloned neurotransmitter receptors to study their pharmacology and
coupling to signal transduction pathways. It is usually assumed that
host cells do not endogenously express the specific receptor under
consideration. We demonstrate in this report that CHO cells contain
endogenous functional muscarinic acetylcholine receptors which, in some
circumstances, might complicate interpretation of data related to the
properties of exogenously expressed receptors.</div></div><br><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MImg&_imagekey=B73F6-47S6466-3F-1&_cdi=11490&_user=108429&_orig=article&_coverDate=10%2F15%2F1995&_sk=997089997&view=c&wchp=dGLbVzW-zSkWA&md5=fe536e47a8f46ddc8c4fb5f772dc2710&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (163 K)</a>
</td></tr></tbody></table></div></td></tr></tbody></table></div><div class="infobubble-container"><div class="mlktLink" id="mlktLink_3"><table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td style="padding: 5px 5px 0px;" valign="top"><img src="science-direct_files/bullet_square.gif" alt=""></td><td onmouseover="InfoBubble.show('infobubble_3','mlktLink_3','iframe_3')" onmouseout="InfoBubble.timeout()"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4JD117M-4&_user=108429&_origUdi=B6TWB-4CGM8MB-1&_fmt=high&_coverDate=07%2F31%2F2006&_rdoc=1&_orig=article&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=8de7af099e2b00eb3d21e264a674bf29">Viability and electrophysiology of neural cell structur...<br><i>Biomaterials</i></a><br><iframe id="iframe_3" style="display: none; position: absolute; z-index: 1;" scrolling="no" frameborder="0"></iframe><div class="infobubble" id="infobubble_3"><table cellpadding="0" cellspacing="0" width="100%"><tbody><tr><td><div style="float: right; padding-left: 5px;"><a onclick="InfoBubble.hide()"><img src="science-direct_files/btn_xclose.gif" alt="Close" title="Close" onmouseover="javascript:this.src='/scidirimg/btn_xclose_hov.gif';" onmouseout="javascript:this.src='/scidirimg/btn_xclose.gif';"></a></div><img src="science-direct_files/jrn_sub.gif" alt="You are entitled to access the full text of this document" title="You are entitled to access the full text of this document" border="0" width="12" height="14"> <b><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4JD117M-4&_user=108429&_coverDate=07%2F31%2F2006&_rdoc=1&_fmt=high&_orig=article&_cdi=5558&_sort=v&_docanchor=&view=c&_ct=1512&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=c9f4ad5625e29646fdaa16fda92968fd">Viability and electrophysiology of neural cell structures generated by the inkjet printing method</a></b><br><i>Biomaterials</i>, <i>Volume 27, Issue 19</i>, <i>July 2006</i>, <i>Pages 3580-3588</i><br>Tao Xu, Cassie A. Gregory, Peter Molnar, Xiaofeng Cui, Sahil Jalota, Sarit B. Bhaduri, Thomas Boland<br><br><b>Abstract</b><br><div class="mlktScroll"><div style="line-height: 150%;">Complex
cellular patterns and structures were created by automated and direct
inkjet printing of primary embryonic hippocampal and cortical neurons.
Immunostaining analysis and whole-cell patch-clamp recordings showed
that embryonic hippocampal and cortical neurons maintained basic
cellular properties and functions, including normal, healthy neuronal
phenotypes and electrophysiological characteristics, after being
printed through thermal inkjet nozzles. In addition, in this study a
new method was developed to create 3D cellular structures: sheets of
neural cells were layered on each other (layer-by-layer process) by
alternate inkjet printing of NT2 cells and fibrin gels. These results
and findings, taken together, show that inkjet printing is rapidly
evolving into a digital fabrication method to build functional neural
structures that may eventually find applications in neural tissue
engineering.</div></div><br><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MImg&_imagekey=B6TWB-4JD117M-4-J&_cdi=5558&_user=108429&_orig=article&_coverDate=07%2F31%2F2006&_sk=999729980&view=c&wchp=dGLbVzW-zSkWA&md5=4a1c82d5beb7200879ee1ca3b56ba8e5&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (504 K)</a>
</td></tr></tbody></table></div></td></tr></tbody></table></div><div class="infobubble-container"><div class="mlktLink" id="mlktLink_4"><table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td style="padding: 5px 5px 0px;" valign="top"><img src="science-direct_files/bullet_square.gif" alt=""></td><td onmouseover="InfoBubble.show('infobubble_4','mlktLink_4','iframe_4')" onmouseout="InfoBubble.timeout()"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6T2C-47MKJKX-BF&_user=108429&_origUdi=B6TWB-4CGM8MB-1&_fmt=high&_coverDate=06%2F30%2F1978&_rdoc=1&_orig=article&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=02c03f254f398aa4d8aa4bcea599e701">Induction of sister<wbr>-<wbr>chromatid exchanges in Chinese hams...<br><i>Mutation Research/Fundamental and Molecular Mechanisms ...</i></a><br><iframe id="iframe_4" style="display: none; position: absolute; z-index: 1;" scrolling="no" frameborder="0"></iframe><div class="infobubble" id="infobubble_4"><table cellpadding="0" cellspacing="0" width="100%"><tbody><tr><td><div style="float: right; padding-left: 5px;"><a onclick="InfoBubble.hide()"><img src="science-direct_files/btn_xclose.gif" alt="Close" title="Close" onmouseover="javascript:this.src='/scidirimg/btn_xclose_hov.gif';" onmouseout="javascript:this.src='/scidirimg/btn_xclose.gif';"></a></div><img src="science-direct_files/jrn_sub.gif" alt="You are entitled to access the full text of this document" title="You are entitled to access the full text of this document" border="0" width="12" height="14"> <b><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6T2C-47MKJKX-BF&_user=108429&_coverDate=06%2F30%2F1978&_rdoc=1&_fmt=high&_orig=article&_cdi=4915&_sort=v&_docanchor=&view=c&_ct=1512&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=0753492349bb11aa70e0eecc3c99b4a5">Induction of sister<wbr>-<wbr>chromatid exchanges in Chinese hamster ovary cells treated in vitro with non<wbr>-<wbr>k<wbr>-<wbr>region dihydrodiols of 7<wbr>-<wbr>methylbenz[<span style="">a</span>]anthracene and benzo[<span style="">a</span>]pyrene</a></b><br><i>Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis</i>, <i>Volume 50, Issue 3</i>, <i>June 1978</i>, <i>Pages 367-375</i><br>Kalyani Pal, Brian Tierney, Philip L. Grover, Peter Sims<br><br><b>Abstract</b><br><div class="mlktScroll"><div style="line-height: 150%;">Studies
were carried out on the incidence of sister-chromatid exchanges induced
in Chinese hamster ovary cells by in vitro treatment with the
polycyclic aromatic hydrocarbons 7-methylbenz[<span style="font-style: italic;">a</span>]anthracene and benzo[<span style="font-style: italic;">a</span>]pyrene
and with related K-region and non-K-region dihydrodiols. Appreciable
increases in the incidence of sister-chromatid exchanges were apparent
in cells treated with non-K-region dihydrodiols: the most active
compounds were 3,4-dihydro-3,4-dihydroxy-7-methylbenz[<span style="font-style: italic;">a</span>]anthracene and 7,8-dihydro-7,8-dihydroxybenzo[<span style="font-style: italic;">a</span>]pyrene
and the effects were dose-dependent. The parent hydrocarbons and the
related K-region dihydrodiols induced some sister-chromatid exchanges
but they were considerably less active than these two non-K-region
diols. The results suggest that this system may usefully be applied to
studies aimed at determining which dihydrodiols are important in the
metabolic activation of the carcinogenic polycyclic hydrocarbons. These
and other results also infer that Chinese hamster ovary cells possess
some intrinsic ability to metabolize such compounds in the absence of
exogenous activation systems.</div></div><br><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MImg&_imagekey=B6T2C-47MKJKX-BF-H&_cdi=4915&_user=108429&_orig=article&_coverDate=06%2F30%2F1978&_sk=999499996&view=c&wchp=dGLbVzW-zSkWA&md5=bdfe8284953baf1bc53d18095bb9d46f&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (555 K)</a>
</td></tr></tbody></table></div></td></tr></tbody></table></div><div class="infobubble-container"><div class="mlktLink" id="mlktLink_5"><table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td style="padding: 5px 5px 0px;" valign="top"><img src="science-direct_files/bullet_square.gif" alt=""></td><td onmouseover="InfoBubble.show('infobubble_5','mlktLink_5','iframe_5')" onmouseout="InfoBubble.timeout()"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4P0N8YT-1&_user=108429&_origUdi=B6TWB-4CGM8MB-1&_fmt=high&_coverDate=09%2F30%2F2007&_rdoc=1&_orig=article&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=d8546a23b309dc0b5f9c1f6170e6486c">Inkjet printing of macromolecules on hydrogels to steer...<br><i>Biomaterials</i></a><br><iframe id="iframe_5" style="display: none; position: absolute; z-index: 1;" scrolling="no" frameborder="0"></iframe><div class="infobubble" id="infobubble_5"><table cellpadding="0" cellspacing="0" width="100%"><tbody><tr><td><div style="float: right; padding-left: 5px;"><a onclick="InfoBubble.hide()"><img src="science-direct_files/btn_xclose.gif" alt="Close" title="Close" onmouseover="javascript:this.src='/scidirimg/btn_xclose_hov.gif';" onmouseout="javascript:this.src='/scidirimg/btn_xclose.gif';"></a></div><img src="science-direct_files/jrn_sub.gif" alt="You are entitled to access the full text of this document" title="You are entitled to access the full text of this document" border="0" width="12" height="14"> <b><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4P0N8YT-1&_user=108429&_coverDate=09%2F30%2F2007&_rdoc=1&_fmt=high&_orig=article&_cdi=5558&_sort=v&_docanchor=&view=c&_ct=1512&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=d8120071ebaa79f092b7fdb62e6bfe86">Inkjet printing of macromolecules on hydrogels to steer neural stem cell differentiation</a></b><br><i>Biomaterials</i>, <i>Volume 28, Issue 27</i>, <i>September 2007</i>, <i>Pages 3936-3943</i><br>Shirin Ilkhanizadeh, Ana I. Teixeira, Ola Hermanson<br><br><b>Abstract</b><br><div class="mlktScroll"><div style="line-height: 150%;">Inkjet
printing allows for the rapid and inexpensive printing of cells,
materials, and protein molecules. However, the combination of inkjet
printing and control of neural stem cell (NSC) multipotency and
differentiation has remained unexplored. We used an inkjet printer
(Canon BJC-2100) to print biologically active macromolecules on
poly-acrylamide-based hydrogels (HydroGel<sup>TM</sup>), which were
subsequently seeded with primary fetal NSCs. NSCs cultured on areas
printed with fibroblast growth factor-2 (FGF2) remained
undifferentiated, consistent with the effects of FGF2 when administered
in solution. NSCs cultured in parallel on the same hydrogels but in
areas printed with ciliary neurotrophic factor (CNTF) or fetal bovine
serum (FBS) displayed a rapid induction of markers for astrocytic
(glial fibrillary acidic protein, GFAP) or smooth muscle (smooth muscle
actin, SMA) differentiation, respectively. These results are consistent
with known actions of CNTF and FBS on NSCs. Importantly, NSCs cultured
on a printed gradient of increasing levels of CNTF showed a linear
increase in numbers of cells expressing GFAP, demonstrating a
functional gradient of CNTF. Lastly, genetically modified NSCs proved
to respond properly to printed macromolecules, suggesting that inkjet
printing can successfully be combined with gene delivery to achieve
effective control of stem cell differentiation.</div></div><br><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MImg&_imagekey=B6TWB-4P0N8YT-1-F&_cdi=5558&_user=108429&_orig=article&_coverDate=09%2F30%2F2007&_sk=999719972&view=c&wchp=dGLbVzW-zSkWA&md5=9e88eaf96d1b877af317f9a1f2c37e62&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (382 K)</a>
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Copyright © 2004 Elsevier Ltd. All rights reserved.<br><br>
<div class="articleTitle">
Inkjet printing of viable mammalian cells </div><!-- articleText -->
<strong>
<p>Tao Xu, Joyce Jin, Cassie Gregory, James J. Hickman<a name="m4.bcor*"></a><a href="#m4.cor*" onclick="toggleTabs('fullTab')"><sup><img src="science-direct_files/REcor.gif" alt="Corresponding Author Contact Information" title="Corresponding Author Contact Information" border="0"></sup></a> and Thomas Boland<a name="m4.bcor*"></a><a href="#m4.cor*" onclick="toggleTabs('fullTab')"><sup><img src="science-direct_files/REcor.gif" alt="Corresponding Author Contact Information" title="Corresponding Author Contact Information" border="0"></sup></a><sup>, </sup><a href="mailto:tboland@clemson.edu"><sup><img src="science-direct_files/REemail.gif" alt="E-mail The Corresponding Author" title="E-mail The Corresponding Author" border="0"></sup></a></p> </strong>
<div style="display: inline;" class="articleText">
<p>Department of Bioengineering, Clemson University, 502 Rhodes Hall, Clemson, SC 29634, USA</p> </div><!-- articleText -->
<div style="display: inline;" class="articleText"><br>
Received 19 December 2003; </div><!-- articleText --><div style="display: inline;" class="articleText">accepted 1 April 2004. </div><!-- articleText --><div style="display: inline;" class="articleText">Available online 28 May 2004. </div><!-- articleText -->
<br><div style="display: inline;" class="articleText"></div><!-- articleText --><div style="display: inline;" class="articleText">
<h3 class="h3">Abstract</h3><p>The purpose of this study was to explore
the use of a commercial thermal printer to deposit Chinese Hamster
Ovary (CHO) and embryonic motoneuron cells into pre-defined patterns.
These experiments were undertaken to verify the biocompatibility of
thermal inkjet printing of mammalian cells and the ability to assemble
them into viable constructs. Using a modified Hewlett Packard (HP) 550C
computer printer and an HP 51626a ink cartridge, CHO cells and rat
embryonic motoneurons were suspended separately in a concentrated
phosphate buffered saline solution (3×). The cells were subsequently
printed as a kind of “ink” onto several “bio-papers” made from soy agar
and collagen gel. The appearance of the CHO cells and motoneurons on
the bio-papers indicated an healthy cell morphology. Furthermore, the
analyses of the CHO cell viability showed that less than 8% of the
cells were lysed during printing. These data indicate that mammalian
cells can be effectively delivered by a modified thermal inkjet printer
onto biological substrates and that they retain their ability to
function. The computer-aided inkjet printing of viable mammalian cells
holds potential for creating living tissue analogs, and may eventually
lead to the construction of engineered human organs.</p> </div><!-- articleText -->
<div style="display: inline;" class="articleText">
<p><strong>Author Keywords: </strong>Inkjet technology; Tissue engineering; Viable mammalian cells</p> </div><!-- articleText -->
<div style="display: inline;" class="articleText">
<h3 class="h3">Article Outline</h3><dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec1">1. Introduction</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec2">2. Materials and methods</a></dt><dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec2.1">2.1. Cell preparation and the cell print suspension (“bio-ink”)</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec2.2">2.2. Bio-paper preparation</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec2.3">2.3. Cell printing and cell culture</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec2.4">2.4. Cell viability</a></dt></dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec3">3. Results</a></dt><dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec3.1">3.1. Cell printing and cell pattern</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec3.2">3.2. CHO cell viability analyses</a></dt></dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec4">4. Discussion</a></dt><dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec4.1">4.1. Cell printing and viability</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec4.2">4.2. Bio-paper</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec4.3">4.3. The cell printer and biocompatibility</a></dt></dl><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#sec5">5. Conclusions</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#ack1">Acknowledgements</a></dt><dt><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bibl1">References</a></dt></dl> </div><!-- articleText -->
<div style="display: inline;" class="articleText">
<a name="sec1"></a><h3 class="h3">1. Introduction</h3><p>This work has
demonstrated that viable mammalian cells be delivered in pre-determined
patterns using a modified inkjet printer. This new ability has
implications for any application that requires spatially registered
cellular engineering, primarily in the field of tissue engineering,
which is a rapidly expanding approach to address the shortage of organs
for transplantation [<a name="bbib1"></a><span class="refPreview" id="refp_1"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib1" onmouseover="RefPreview.showRef(event,'ref_bib1','refp_1')" onmouseout="RefPreview.hideRef()">1</a>]. In the most general sense, tissue engineering seeks to fabricate viable replacement parts for the body [<a name="bbib2"></a><span class="refPreview" id="refp_2"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib2" onmouseover="RefPreview.showRef(event,'ref_bib2','refp_2')" onmouseout="RefPreview.hideRef()">2</a>].
Although still in its infancy, much progress in tissue engineering has
been made in areas relevant to the development of novel biomaterials [<a name="bbib3"></a><span class="refPreview" id="refp_3"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib3" onmouseover="RefPreview.showRef(event,'ref_bib3','refp_3')" onmouseout="RefPreview.hideRef()">3</a>], the design of bioreactors for dynamic in vitro culture systems [<a name="bbib4"></a><span class="refPreview" id="refp_4"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib4" onmouseover="RefPreview.showRef(event,'ref_bib4','refp_4')" onmouseout="RefPreview.hideRef()">4</a>], and the use of a wide variety of cell sources, including appropriate multi-potent stem cells [<a name="bbib5"></a><span class="refPreview" id="refp_5"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib5" onmouseover="RefPreview.showRef(event,'ref_bib5','refp_5')" onmouseout="RefPreview.hideRef()">5</a>].
Advancements in tissue engineering have permitted the creation of
functional tissue substitutes available to patients for clinical
applications, including engineered stomachs [<a name="bbib6"></a><span class="refPreview" id="refp_6"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib6" onmouseover="RefPreview.showRef(event,'ref_bib6','refp_6')" onmouseout="RefPreview.hideRef()">6</a>], esophagus [<a name="bbib7"></a><span class="refPreview" id="refp_7"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib7" onmouseover="RefPreview.showRef(event,'ref_bib7','refp_7')" onmouseout="RefPreview.hideRef()">7</a>], spinal cord [<a name="bbib8"></a><span class="refPreview" id="refp_8"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib8" onmouseover="RefPreview.showRef(event,'ref_bib8','refp_8')" onmouseout="RefPreview.hideRef()">8</a>], and progress towards complete tissue engineered organs [<a name="bbib9"></a><span class="refPreview" id="refp_9"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib9" onmouseover="RefPreview.showRef(event,'ref_bib9','refp_9')" onmouseout="RefPreview.hideRef()">9</a>].
To further the construction of complex tissues, or even entire organs,
however, there are still significant technical challenges to overcome.
Among them, a very important problem is subtly combining and
orchestrating cells, growth factors and scaffolds into an architecture
that will allow their unfettered interaction, especially where distinct
cell types are required in anatomically exact locations to attain
biological function [<a name="bbib10"></a><span class="refPreview" id="refp_10"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib10" onmouseover="RefPreview.showRef(event,'ref_bib10','refp_10')" onmouseout="RefPreview.hideRef()">10</a>]. Inkjet printing technology offers a possible solution to this complex problem.</p><p>Inkjet printing, initially from the field of electronics and mechanics [<a name="bbib11"></a><span class="refPreview" id="refp_11"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib11" onmouseover="RefPreview.showRef(event,'ref_bib11','refp_11')" onmouseout="RefPreview.hideRef()">11</a>], has recently been extended to bioengineering applications [<a name="bbib12"></a><span class="refPreview" id="refp_12"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib12" onmouseover="RefPreview.showRef(event,'ref_bib12','refp_12')" onmouseout="RefPreview.hideRef()">12</a>].
With the obvious advantages of being inexpensive as well as high
throughput, commercial thermal inkjet printers have been modified to
print biomolecules onto target substrates with little or no reduction
of their bioactivities, resulting in the creation of DNA chips [<a name="bbib13"></a><span class="refPreview" id="refp_13"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib13" onmouseover="RefPreview.showRef(event,'ref_bib13','refp_13')" onmouseout="RefPreview.hideRef()">13</a>], protein arrays [<a name="bbib14"></a><span class="refPreview" id="refp_14"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib14" onmouseover="RefPreview.showRef(event,'ref_bib14','refp_14')" onmouseout="RefPreview.hideRef()">14</a>] and cell patterns [<a name="bbib15"></a><span class="refPreview" id="refp_15"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib15" onmouseover="RefPreview.showRef(event,'ref_bib15','refp_15')" onmouseout="RefPreview.hideRef()">15</a>]. In particular, our recent success in printing viable bacteria directly using an off-the-self thermal inkjet printer [<a name="bbib16"></a><span class="refPreview" id="refp_16"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib16" onmouseover="RefPreview.showRef(event,'ref_bib16','refp_16')" onmouseout="RefPreview.hideRef()">16</a>]
significantly expands the capabilities of inkjet printing in the
direction of creating living tissue substitutes. By means of
computer-assisted deposition, viable cells can be delivered to precise
target positions on scaffold materials. Furthermore, using different
cell types as different bio-inks, which are then delivered to exact
positions to mimic tissue structures of the original tissue, can be
envisioned by using multiple nozzles. Thus, the printing of dissociated
human or animal cells onto specific patterns, and their subsequent
fusion, may allow the development of replacement tissue or even whole
organ substitutes [<a name="bbib17"></a><span class="refPreview" id="refp_17"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib17" onmouseover="RefPreview.showRef(event,'ref_bib17','refp_17')" onmouseout="RefPreview.hideRef()">17</a>].
Compared to bacteria, however, animal cells are generally more
sensitive to heat and mechanical stress, both of which often occur in
the cartridge of a printer. Whether animal cells could be effectively
printed using the modified thermal printer still needed to be
demonstrated to realize this ambitious goal.</p><p>In the present
study, suspensions of Chinese Hamster Ovary (CHO) and embryonic rat
motoneurons were printed directly using a modified Hewlett Packard (HP)
550C printer. This thermal printer was chosen over the piezo-electric
technology because our previous experience in adapting these commercial
inkjet printers for biomaterial printing [<span class="refPreview" id="refp_18"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib15" onmouseover="RefPreview.showRef(event,'ref_bib15','refp_18')" onmouseout="RefPreview.hideRef()">15</a>, <span class="refPreview" id="refp_19"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib16" onmouseover="RefPreview.showRef(event,'ref_bib16','refp_19')" onmouseout="RefPreview.hideRef()">16</a>, <span class="refPreview" id="refp_20"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib17" onmouseover="RefPreview.showRef(event,'ref_bib17','refp_20')" onmouseout="RefPreview.hideRef()">17</a>, <a name="bbib18"></a><span class="refPreview" id="refp_21"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib18" onmouseover="RefPreview.showRef(event,'ref_bib18','refp_21')" onmouseout="RefPreview.hideRef()">18</a> and <a name="bbib19"></a><span class="refPreview" id="refp_22"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib19" onmouseover="RefPreview.showRef(event,'ref_bib19','refp_22')" onmouseout="RefPreview.hideRef()">19</a>].
Soy agar and collagen hydrogels were used as the culture substrate or
“bio-paper” to prevent the cells from drying and to provide nutrients
for cell growth. The cell viability during the printing was evaluated
by a combined lactate dehydrogenase (LDH) and tetrazolium compound
assay (MTS).</p><h3 class="h3"><a name="sec2"></a>2. Materials and methods</h3><a name="sec2.1"></a><h4 class="h4">2.1. Cell preparation and the cell print suspension (“bio-ink”)</h4><p>Two
different cell types were used for the inkjet printing experiments. One
was a cell line of Chinese Hamster Ovary (CHO-B2) cells. These cells
had been transfected to express green fluorescent protein and <i>α</i>5<i>β</i>1 integrin which causes them to form compact, spherical aggregates [<a name="bbib20"></a><span class="refPreview" id="refp_23"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib20" onmouseover="RefPreview.showRef(event,'ref_bib20','refp_23')" onmouseout="RefPreview.hideRef()">20</a>].
Cells with green fluorescence are easy to observe for the printed
patterns and other applications. The other cells used in this study
were primary embryonic motoneurons obtained from ventral cords of day
14 embryos from pregnant Sprague–Dawley rats as previously described [<a name="bbib21"></a><span class="refPreview" id="refp_24"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib21" onmouseover="RefPreview.showRef(event,'ref_bib21','refp_24')" onmouseout="RefPreview.hideRef()">21</a>].</p><p>Cell
pellets were collected by centrifugation (1000 rpm/2 min) in
a conical tube. After aspirating the supernatant, they were resuspended
in 0.5 ml of 3× Dulbecco's phosphate buffered saline solution
(DPBS) (Sigma Chemical Inc., St. Louis, MO) to obtain cell print
suspensions (like “inks” for the printers). The concentration of the
CHO cell's print suspension was about 5,000,000 cells/ml, and the
primary motoneuron concentration was around 2,000,000 cells/ml.</p><a name="sec2.2"></a><h4 class="h4">2.2. Bio-paper preparation</h4><p>The
two hydrogel-based substrates were prepared as the print substrates (or
bio-papers). One of the bio-papers used was soy agar gel, and the other
was a collagen gel. Both were coated on glass coverslips (22×22 mm<sup>2</sup>). The soy agar gel was made from Trypticase<sup>®</sup>
soy agar solution (Becton Dickinson & Co., Cockeysville, MD) by
dissolving 1.5 g of powdered soy agar in 50 ml of nanopure
water. After autoclaving for 15 min, 50 ml of a 2× solution
of Dulbecco's modified Eagles medium (DMEM) (Sigma) was added to the
soy agar solution while it still was relatively hot (about 45°C). After
mixing thoroughly, 2 ml aliquots of the mixture were dispensed
onto each coverslip, which was in 35 mm dishes. The soy agar gel
formed on the coverslips after cooling to room temperature. The
collagen gel bio-paper was produced from rat-tail Type I collagen (BD
Biosciences, Billerica, MA), which was received at a concentration of
3.67 mg/ml in acetic acid. The collagen solution obtained a final
concentration of 1.0 mg/ml by adding chilled PBS (2×). After
adjusting the pH to about 7.4, 1.5 ml aliquots of the mixture were
dispersed onto coverslips and plated in an incubator for 3–5 h.
Once the gel formed, the bio-paper of collagen was ready for use.</p><a name="sec2.3"></a><h4 class="h4">2.3. Cell printing and cell culture</h4><p>A
modified HP Desktop printer (HP 550C) and a modified HP 51626a ink
cartridge were used to print the CHO cells and motoneurons directly
into specific patterns. The printer modifications are described
previously [<span class="refPreview" id="refp_25"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib18" onmouseover="RefPreview.showRef(event,'ref_bib18','refp_25')" onmouseout="RefPreview.hideRef()">18</a>].
For these experiments, after emptying the ink, the cartridge was rinsed
thoroughly with 100% ethanol and sterile water. Previously, this
procedure was proven to be enough to create uncontaminated bacterial
cell cultures [<span class="refPreview" id="refp_26"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib16" onmouseover="RefPreview.showRef(event,'ref_bib16','refp_26')" onmouseout="RefPreview.hideRef()">16</a>].
A pattern that consisted of rows of circles was designed using
Microsoft PowerPoint software (Microsoft, Redmond, WA) to program the
printer.</p><p>To form the patterns, the cell print suspension was
shaken forcefully, and then a small portion (0.3–0.5 ml) of the
suspension was loaded into the ink-containing chamber on the bottom of
the cartridge. The hydrogel-based bio-papers were placed onto the print
basement of the modified printer. The cells were delivered onto the
bio-paper following the designed pattern and the cells were printed
several consecutive times (2–8×) at the same position to ensure a high
cell number on the circular patterns. After printing, the bio-papers
with the printed cells were placed into dishes and moved to the
incubator, which was maintained at 37°C, 5% CO<sub>2</sub>, 95% air and
99% relative humidity. After 1–2 h incubation, a small amount of
culture medium was carefully added to the dishes to avoid disturbing
the printed cell patterns and the level of the culture medium was
maintained to just below the top layer of the gel. DMEM medium,
supplemented with 10% fetal bovine serum (Sigma), 0.15% (w/v) sodium
bicarbonate and 1% antibiotic/antimycotic (Sigma Chemical Inc.)
solution was used to feed the CHO cells. The medium was changed every
1–2 days and the growth of the cells was monitored daily via light and
epifluorescent microscopy. The culture medium for the primary
motoneurons was Neurobasal (Gibco-BRL) supplemented with B27 (2% v/v)
(Invitrogen, Carlsbad, CA), <img src="science-direct_files/0_004.gif" alt="Image" title="Image" align="absbottom" border="0" width="9" height="10">
-glutamine (0.5 m<img src="science-direct_files/0_003.gif" alt="Image" title="Image" align="absbottom" border="0" width="13" height="10">
) and 2-mercaptoethanol (25 μ<img src="science-direct_files/0_003.gif" alt="Image" title="Image" align="absbottom" border="0" width="13" height="10">
). The culture medium was changed every four days and <img src="science-direct_files/0_004.gif" alt="Image" title="Image" align="absbottom" border="0" width="9" height="10">
-glutamate (25 μ<img src="science-direct_files/0_003.gif" alt="Image" title="Image" align="absbottom" border="0" width="13" height="10">
) was added to the medium during the first four days of culture.</p><a name="sec2.4"></a><h4 class="h4">2.4. Cell viability</h4><p>The
amount of LDH, a stable cytosolic enzyme that is released upon cell
lysis, was measured quantitatively using a commercial calorimetric
assay (Cytotox 96, Promega Corp., Madison, WI). Per the manufacturer's
protocol, 500 μl of reagent was added per ml of media into culture
dishes containing the printed samples. As a control, a known number of
cells (0, 1000, 5000, 10000, and 20,000, as determined by triplicate
hemocytometer counts) were lysed using 9% (v/v) Triton<sup>®</sup>
X-100; the other controls were samples coated with the gel but without
cells. The absorbance at 490 nm of the solution was measured after
30 min of incubation at room temperature in the dark. The amount
of live cells was determined by a tetrazolium compound assay (MTS;
CellTiter 96 Promega Corp., Madison, WI). Per the manufacturer's
protocol, 500 μl of reagent was added per ml of media into the
culture dishes previously used for the LDH assay. The absorbances for a
known amount of cells as well as for the pure gel samples were used as
controls. The samples and controls were incubated for 60 min at
37°C in the dark. The total number of cells was calculated as the sum
of the lysed and metabolically active cells, and the percentage of
cells lysed by printing was estimated as the ratio of the lysed cells
to the total number of cells.</p><h3 class="h3"><a name="sec3"></a>3. Results</h3><a name="sec3.1"></a><h4 class="h4">3.1. Cell printing and cell pattern</h4><p>The
CHO cells were observed to still emit green fluorescence by
epifluorescent microscopy, which strongly suggested that the CHO cells
were still living after passing through the nozzle of the cartridge.
During the initial 5 days of culture, the printed cells did not from a
complete ring because the initial plating was at low density, which
obscured the ring pattern. After 6 days, the cells gradually occupied
the positions along the ring pattern to form parts of the cell ring,
indicating the retention of the ability to proliferate and the
long-term viability of the technique. At day 7, the complete ring of
CHO cells was observed on the soy agar bio-paper (<span class="refPreview" id="refp_27"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#fig1" onmouseover="RefPreview.showRef(event,'ref_fig1','refp_27')" onmouseout="RefPreview.hideRef()">Fig. 1A</a>).
The cells continued to proliferate until a complete circle was formed
by day 25. Since CHO cells are not contact inhibited, eventually the
entire gel became populated with cells.</p></div><div class="figTblUpiOuter"><div style="display: inline;" class="graphText"><br><br><a name="fig1"></a><a name="1"></a><table><tbody><tr><td width="10%"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig1&_ba=1&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=ec20d0cf98a97912391d465ad9ec6ea1" onclick="openStrippedNS('/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig1&_ba=1&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=ec20d0cf98a97912391d465ad9ec6ea1'); return false;"><img src="science-direct_files/0.gif" title="Full-size image (89K) - Opens new window" alt="Full-size image (89K) - Opens new window" align="middle" border="1" vspace="2" width="90" height="93" hspace="2"></a></td><td valign="bottom"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig1&_ba=1&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=ec20d0cf98a97912391d465ad9ec6ea1" onclick="openStrippedNS('/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig1&_ba=1&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=ec20d0cf98a97912391d465ad9ec6ea1'); return false;" alt="Full-size image (89K) - Opens new window" title="Full-size image (89K) - Opens new window">Full-size image</a> (89K)</td></tr></tbody></table><p>Fig.
1. Chinese hamster ovary cells visualized under a fluorescent
microscope. The pattern as seen on the agar gel bio-paper at 7 days of
culture (A); the pattern after 15 days of culture (B); the pattern
after 20 days of culture (C), and the pattern after 25 days of culture
(D). Since CHO cells are not contact-inhibited, the cells eventually
grew everywhere in the gel bio-paper. Magnifications are as follows: 2×
(A), and 12× (B, C, and D).</p> <div class="graphTextOnly" style="display: none;"><br><a href="#fig1" onclick="toggleTabs('fullTab');" onmouseover="javascript:this.style.cursor='pointer'"><div class="viewWithinBtn" style="background-image: url(/scidirimg/midbg.gif);">View Within Article</div><!--viewwithinbtn--></a><br></div><!--graphTextOnly--></div><!--graphText--></div><div style="display: inline;" class="articleText"><br><p>A printed neuronal ring was obvious after 1 day in culture, as shown in <span class="refPreview" id="refp_28"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#fig2" onmouseover="RefPreview.showRef(event,'ref_fig2','refp_28')" onmouseout="RefPreview.hideRef()">Fig. 2A</a>.
Many rounded cells were seen, indicative of non-differentiated or dead
cells. Most of the cells that survived the passage through the nozzle
grew processes. Few differentiated neurons with processes were obvious
at the pattern at day 2 in culture, but after 5 days, many neurons
began to differentiate and developed processes, exhibiting a polarized
morphology (<span class="refPreview" id="refp_29"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#fig2" onmouseover="RefPreview.showRef(event,'ref_fig2','refp_29')" onmouseout="RefPreview.hideRef()">Figs. 2C and D</a>).</p></div><div class="figTblUpiOuter"><div style="display: inline;" class="graphText"><div class="graphTextOnly" style="display: none;"><hr><br></div><br><br><a name="fig2"></a><a name="2"></a><table><tbody><tr><td width="10%"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig2&_ba=2&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f98088b605fa55be5788cd1971af8f08" onclick="openStrippedNS('/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig2&_ba=2&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f98088b605fa55be5788cd1971af8f08'); return false;"><img src="science-direct_files/0_002.gif" title="Full-size image (92K) - Opens new window" alt="Full-size image (92K) - Opens new window" align="middle" border="1" vspace="2" width="77" height="93" hspace="2"></a></td><td valign="bottom"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig2&_ba=2&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f98088b605fa55be5788cd1971af8f08" onclick="openStrippedNS('/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=fig2&_ba=2&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f98088b605fa55be5788cd1971af8f08'); return false;" alt="Full-size image (92K) - Opens new window" title="Full-size image (92K) - Opens new window">Full-size image</a> (92K)</td></tr></tbody></table><p>Fig.
2. Primary motoneurons were delivered by direct inkjet printing in the
pre-designed ring patterns as indicated as at day 1 utilizing bright
field miscopy (A). Motoneurons on the ring patterns with developing
processes at day 2 observed with bright field miscopy. A few neurons
are extending their processes (B). At day 3 many of the motoneurons
exhibited a characteristic neuronal morphology as seen with bright
field miscopy (C). At day 7 the processes of two neurons have grown to
about 40 microns in length (D). Magnifications are as follows: 40× (A),
100× (B and C), and 400× (D).</p> <div class="graphTextOnly" style="display: none;"><br><a href="#fig2" onclick="toggleTabs('fullTab');" onmouseover="javascript:this.style.cursor='pointer'"><div class="viewWithinBtn" style="background-image: url(/scidirimg/midbg.gif);">View Within Article</div><!--viewwithinbtn--></a><br></div><!--graphTextOnly--></div><!--graphText--></div><div style="display: inline;" class="articleText"><br><a name="sec3.2"></a><h4 class="h4">3.2. CHO cell viability analyses</h4><p><span class="refPreview" id="refp_30"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#tbl1" onmouseover="RefPreview.showRef(event,'ref_tbl1','refp_30')" onmouseout="RefPreview.hideRef()">Table 1</a>
shows the percentages of lysed CHO cells after the bio-ink formulation
and the printing process. On average, a very small amount of cells was
lysed during the printing process, 3.3±3.7% (<i>n</i>=4); but in all
cases the number of lysed cells was below 10%. The total number of
printed cells ranged from 44,000 to 154,000 in these experiments. The
number depended on the particular pattern and the number of repetitive
nozzle firings. These numbers are well below the manual controls, which
may explain the high standard deviations. The bio-ink formulation lysed
on average 15±0.7% of the CHO cells.</p></div><div class="figTblUpiOuter"><div style="display: inline;" class="graphText"><div class="graphTextOnly" style="display: none;"><hr><br></div><a name="tbl1"></a><p>Table
1. Bio-ink formulation and printing cytotoxicity as indicated by the
combined lactate dehydrogenase (LDH) and tetrazolium compound assay
(MTS)</p> <div class="tblScrollBox"><div class="tblScroll"><img src="science-direct_files/0_005.gif" alt="Image" title="Image" align="absbottom" border="0" width="767" height="136">
<br></div><!--tblScroll--><div style="display: none;" class="tblScrollLink"><a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=tbl1&_ba=&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_isTablePopup=Y&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=5386d1c9cae18e5965adb626e893cf13" onclick="openStrippedNS('/science?_ob=MiamiCaptionURL&_method=retrieve&_udi=B6TWB-4CGM8MB-1&_image=tbl1&_ba=&_user=10&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&view=c&_isTablePopup=Y&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=5386d1c9cae18e5965adb626e893cf13'); return false;" alt="Full-size table (9K) - Opens new window" title="Full-size table (9K) - Opens new window">Full-size table</a> (9K)</div><!--tblScrollLink--></div><!--tblScrollBox--><div class="graphTextOnly" style="display: none;"><br><a href="#tbl1" onclick="toggleTabs('fullTab');" onmouseover="javascript:this.style.cursor='pointer'"><div class="viewWithinBtn" style="background-image: url(/scidirimg/midbg.gif);">View Within Article</div><!--ViewWithinBtn--></a><br></div><!--graphTextOnly--></div><!--graphText--></div><div style="display: inline;" class="articleText"><h3 class="h3"><a name="sec4"></a>4. Discussion</h3><a name="sec4.1"></a><h4 class="h4">4.1. Cell printing and viability</h4><p>By
replacing the regular print ink, the cell suspensions can be considered
“bio-ink” that could be delivered using a commercial, thermal-based
inkjet printer. Our previous study proved the ability to print viable
bacteria directly by the inkjet printing method [<span class="refPreview" id="refp_31"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib16" onmouseover="RefPreview.showRef(event,'ref_bib16','refp_31')" onmouseout="RefPreview.hideRef()">16</a>].
However, it had not been reported whether viable mammalian cells, the
cells needed for building engineered tissues and organs, could be used
as a bio-ink and printed directly by a commercial inkjet printer. Since
the physiological properties of mammalian cells strongly depend on the
culture conditions and they are much more sensitive to heat and
mechanical stress, there was a major concern that the cells could be
damaged or lysed by the conditions present during thermal printing. The
temperature in the nozzle of the cartridge can be 300°C or higher [<a name="bbib22"></a><span class="refPreview" id="refp_32"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib22" onmouseover="RefPreview.showRef(event,'ref_bib22','refp_32')" onmouseout="RefPreview.hideRef()">22</a>].
The present study indicates, however, that viable CHO cells and primary
motoneurons can be delivered successfully by using a modified inkjet HP
printer, and most of these cells (>90%) were not lysed during
printing as shown in <span class="refPreview" id="refp_33"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#tbl1" onmouseover="RefPreview.showRef(event,'ref_tbl1','refp_33')" onmouseout="RefPreview.hideRef()">Table 1</a>.</p><p>These data confirm our previous analysis, that cells can survive thermal printing under low power conditions [<span class="refPreview" id="refp_34"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib16" onmouseover="RefPreview.showRef(event,'ref_bib16','refp_34')" onmouseout="RefPreview.hideRef()">16</a>].
The HP inkjet printer technology is based on vaporizing a
micrometer-sized layer of liquid in contact with a thin film resistor.
As the timescales involved in the drop ejection process are small,
there is not enough time for heat to diffuse into the bulk liquid.
While the surface in contact with the liquid can peak at 250–350°C, the
bulk liquid does not rise more than a couple (approximately 4–10°C)
degrees above ambient [<a name="bbib23"></a><span class="refPreview" id="refp_35"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib23" onmouseover="RefPreview.showRef(event,'ref_bib23','refp_35')" onmouseout="RefPreview.hideRef()">23</a>].
This situation could change however, depending on the liquid
thermophysical properties of the ink. Although mammalian cells are more
sensitive to high heat and strong mechanical stress than bacteria,
their volume is also relatively small compared to the total volume of
the printed droplet. The fact that these mammalian cells are viable,
can proliferate and differentiate indicates that damage from the heat
and mechanical stress during the very short timescale of printing is
avoided. Moreover, the 3× PBS used for the cell print suspensions could
have caused a further decrease in cell volume, because they are
expected to shrink by osmosis in this hypertonic solution, effectively
preventing clogging of the nozzle during cell printing. This would come
at the expense of higher cell death in the ink formulation. However,
once deposited on the gel, the CHO cells returned to their normal shape
and morphology to a great extent. The survival of motoneurons was
indicated by the outgrowth of their processes and the establishment of
polarized morphologies ( <span class="refPreview" id="refp_36"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#fig2" onmouseover="RefPreview.showRef(event,'ref_fig2','refp_36')" onmouseout="RefPreview.hideRef()">Figs. 2B–D</a>). However, the lengths of the processes were shorter than that observed in the standard 2D culture [<span class="refPreview" id="refp_37"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib21" onmouseover="RefPreview.showRef(event,'ref_bib21','refp_37')" onmouseout="RefPreview.hideRef()">21</a>].
One of the possible reasons could be related to the use of the 3D
collagen gel based bio-paper. Prior studies have found that increasing
collagen concentration will decrease the neurite outgrowth [<a name="bbib24"></a><span class="refPreview" id="refp_38"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib24" onmouseover="RefPreview.showRef(event,'ref_bib24','refp_38')" onmouseout="RefPreview.hideRef()">24</a>].</p><p>Inkjet
printers based on piezo-electric technology may also be modified for
cell printing as they have many applications in ceramics and polymer
printing. However, there are some challenges in adapting commercial
piezo-printers for printing mammalian cells. For many reasons,
including minimizing ink leakage and preventing formation of mist,
commercial piezo-printers use a more viscous ink. This helps eliminate
the need for complex fluid gates between the ink cartridge and
printhead to prevent the ink from backflowing. However, this comes at
the expense of requiring more power and higher vibration frequencies,
both of which can break and damage the cell membranes. Typical
commercial piezo-printers use frequencies up to 30 kHz and power
sources ranging from 12 to 100 W. This becomes a problem because
vibrating frequencies ranging from 15 to 25 kHz and power sources
from 10 to 375 W are often used to disrupt cell membranes [<a name="bbib25"></a><span class="refPreview" id="refp_39"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib25" onmouseover="RefPreview.showRef(event,'ref_bib25','refp_39')" onmouseout="RefPreview.hideRef()">25</a>].
Adapting piezo-printers for less viscous ink to lower the frequency and
power would be challenging, since ink leakage and mist formation during
printing could obscure the pattern. While these problems may be
overcome, this study focused on adaptation of commercial printers
without significant redesigning.</p><a name="sec4.2"></a><h4 class="h4">4.2. Bio-paper</h4><p>Cell
printing is different from printing of bioactive molecules such as DNA
or proteins. While the latter are deposited onto dry substrates, and
thus dry out quickly after deposition, the former require suitable
materials to prevent drying of the cells. Viability of the cells is
strongly affected by the drying process of the ink droplets after
printing. We have earlier pointed to hydrogels as a possible solution
to the drying problem [<span class="refPreview" id="refp_40"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib19" onmouseover="RefPreview.showRef(event,'ref_bib19','refp_40')" onmouseout="RefPreview.hideRef()">19</a>]
as they can provide the cells with water as well as nutrients and
mechanical support during this time. Two different hydrogels were used
as substrates in this study. For the fast, anchorage independent
proliferating CHO cells, a gel that limits the migration was needed;
thus soy agar hydrogels were used. With no or limited cell motility,
CHO cells survived and multiplied only around the positions where they
were printed, thus allowing the CHO cell pattern to fuse into to a
clear ring rather than a disk. For the anchorage dependent motoneurons,
a gel that promoted differentiation was needed. Soy agar gel could not
be used as a substrate for neuronal printing and pattering as neurons
are thought to be difficult to differentiate on the soy agar gels [<a name="bbib26"></a><span class="refPreview" id="refp_41"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib26" onmouseover="RefPreview.showRef(event,'ref_bib26','refp_41')" onmouseout="RefPreview.hideRef()">26</a>]. Since collagen had been shown to promote neuron differentiation [<a name="bbib27"></a><span class="refPreview" id="refp_42"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib27" onmouseover="RefPreview.showRef(event,'ref_bib27','refp_42')" onmouseout="RefPreview.hideRef()">27</a>],
the collagen gel-based bio-paper was used for the neuronal printing.
These two gels were chosen because they ensured cell survival, division
and differentiation for cells representing either less or fully
differentiated phenotypes.</p><p>The behaviors of cells on or within the bio-paper are directed by cell–material interactions [<span class="refPreview" id="refp_43"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib3" onmouseover="RefPreview.showRef(event,'ref_bib3','refp_43')" onmouseout="RefPreview.hideRef()">3</a> and <a name="bbib28"></a><span class="refPreview" id="refp_44"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib28" onmouseover="RefPreview.showRef(event,'ref_bib28','refp_44')" onmouseout="RefPreview.hideRef()">28</a>].
Recently, it was demonstrated that the self-assembling and/or fusion of
CHO aggregates embedded into 3D hydrogels depended greatly on the
magnitude of cell–cell and cell–matrix interactions in the 3D systems [<span class="refPreview" id="refp_45"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib17" onmouseover="RefPreview.showRef(event,'ref_bib17','refp_45')" onmouseout="RefPreview.hideRef()">17</a> and <a name="bbib29"></a><span class="refPreview" id="refp_46"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib29" onmouseover="RefPreview.showRef(event,'ref_bib29','refp_46')" onmouseout="RefPreview.hideRef()">29</a>].
It was found, depending on the amount of cell adhesive ligands in the
gel, that CHO aggregates either fused into a torodial 3D structure
(when using RGD containing NeuroGel or 1.0 mg/ml collagen gel) or
dispersed into the surrounding matrix (when using 1.7 mg/ml
collagen gel). These findings could lead to the creation of functional
bio-papers, especially when combined with our previous report of
collagen printing [<span class="refPreview" id="refp_47"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib15" onmouseover="RefPreview.showRef(event,'ref_bib15','refp_47')" onmouseout="RefPreview.hideRef()">15</a>].
Collagen, RGD, or other cell-adhesion molecules could be printed first
using a separate nozzle before delivering the cells as a second step to
achieve this goal. Such multiprocessing would combine functional
bio-papers for controlled cell–gel interactions, with automated
delivery of cells, resulting in self-organization of cells or cell
aggregates into metastable tissue structures of the desired shape.</p><a name="sec4.3"></a><h4 class="h4">4.3. The cell printer and biocompatibility</h4><p>Few
tools have been applied effectively to date to directly deliver viable
cells onto pre-defined patterns for the purposes of cell patterning and
tissue construction [<a name="bbib30"></a><span class="refPreview" id="refp_48"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib30" onmouseover="RefPreview.showRef(event,'ref_bib30','refp_48')" onmouseout="RefPreview.hideRef()">30</a>]. The laser-directed cell writing system is one of the few examples of direct cell placement [<a name="bbib31"></a><span class="refPreview" id="refp_49"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib31" onmouseover="RefPreview.showRef(event,'ref_bib31','refp_49')" onmouseout="RefPreview.hideRef()">31</a>].
Limited throughput and the difficulties in extending this technology
into 3D constructs, however, have limited its potential application
into product development or tissue construction. By definition a
non-contact technique [<a name="bbib32"></a><span class="refPreview" id="refp_50"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib32" onmouseover="RefPreview.showRef(event,'ref_bib32','refp_50')" onmouseout="RefPreview.hideRef()">32</a>],
inkjet printers combine micro-electronic, micro-mechanic, and computer
elements into a simple, yet efficient platform, offering a new a
paradigm for the precise delivery of bio-ink droplets onto exact
positions on a substrate. Inspired by traditional inkjet technology,
bio-printers have been developed to perform the computer-assisted
deposition of natural materials, including bioactive molecules [<span class="refPreview" id="refp_51"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib18" onmouseover="RefPreview.showRef(event,'ref_bib18','refp_51')" onmouseout="RefPreview.hideRef()">18</a>], biomolecules [<span class="refPreview" id="refp_52"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib13" onmouseover="RefPreview.showRef(event,'ref_bib13','refp_52')" onmouseout="RefPreview.hideRef()">13</a>, <span class="refPreview" id="refp_53"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib14" onmouseover="RefPreview.showRef(event,'ref_bib14','refp_53')" onmouseout="RefPreview.hideRef()">14</a> and <span class="refPreview" id="refp_54"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib15" onmouseover="RefPreview.showRef(event,'ref_bib15','refp_54')" onmouseout="RefPreview.hideRef()">15</a>] and cells [<span class="refPreview" id="refp_55"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib16" onmouseover="RefPreview.showRef(event,'ref_bib16','refp_55')" onmouseout="RefPreview.hideRef()">16</a> and <span class="refPreview" id="refp_56"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib19" onmouseover="RefPreview.showRef(event,'ref_bib19','refp_56')" onmouseout="RefPreview.hideRef()">19</a>].
Direct patterning of viable mammalian cells onto hydrogel-bases papers
using our cell printer could be a significant advance in living
biomaterial processing and shows progress in our understanding and
manipulation of natural systems. The cell printer juxtaposes
electronic, mechanic, and biomaterial platforms, and is compatible with
all key elements required for precise arrangement of viable cell in
tissue engineering, as well as those of the control of biological
molecules [<span class="refPreview" id="refp_57"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib13" onmouseover="RefPreview.showRef(event,'ref_bib13','refp_57')" onmouseout="RefPreview.hideRef()">13</a>].
Moreover, this cell printer provides a useful tool for building 3D
living tissues or even organ substitutes, especially when used in
combination with the ability to print other cell-friendly materials or
scaffolds adjacent to viable cells, as shown by our previous work to
rapidly fabricate cell patterns [<span class="refPreview" id="refp_58"></span><a onclick="toggleTabs('fullTab')" href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6TWB-4CGM8MB-1&_user=108429&_coverDate=01%2F31%2F2005&_rdoc=1&_fmt=full&_orig=search&_cdi=5558&_sort=d&_docanchor=&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=f74163947b3d8c9759985d09f284b7c9#bib15" onmouseover="RefPreview.showRef(event,'ref_bib15','refp_58')" onmouseout="RefPreview.hideRef()">15</a>].
While this initial study was mainly concerned with the survivability of
mammalian cells, future studies will need to optimize bio-ink and
bio-paper formulations for this technology to be successfully used in
the field of tissue engineering.</p><a name="sec5"></a><h3 class="h3">5. Conclusions</h3><p>The
CHO and motoneuron cells were successfully printed through the nozzles
of a thermal inkjet printer. The printed cells were deposited onto
gel-based bio-papers with different adhesive properties to form
pre-defined patterns. The analysis of the CHO cell viability showed
that more than 90% of cells were not damaged during the nozzle firing.
An exciting combination of off-the-shelf inkjet printing and viable
biomaterials was demonstrated for cell patterning and now could be
adapted to the application of tissue engineering. With the obvious
advantages of high throughput and flexibility, this bottom-up
technology offers researchers a cost-effective tool to rapidly
fabricate cell patterns and tissue-like structures.</p> </div><!-- articleText -->
<div style="display: inline;" class="articleText">
<a name="ack1"></a><h3 class="h3">Acknowledgements </h3><p>The authors
would like to acknowledge other members of the Clemson Bioengineering
Department whose help has made this study possible. We would thank the
Hunter Chair Endowment, IPTO office of DARPA (grant #
N65236-01-1-7400), and the Department of Energy (grant #
DE-FG02-00ER-45856) for funding this project.</p> </div><!-- articleText -->
<div style="display: inline;" class="refText">
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| <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=655&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Frecord.url%3Feid%3D2-s2.0-0037408143%26partnerID%3D10%26rel%3DR3.0.0%26md5%3D9db73034932a9fd8eb7840b66e4671ba&_acct=C000059713&_version=1&_userid=108429&md5=be5b26d2638cc2d9d2438c7ee740352e" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">View Record in Scopus</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=656&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Fcitedby.url%3Feid%3D2-s2.0-0037408143%26partnerID%3D10%26rel%3DR3.0.0%26md5%3D9db73034932a9fd8eb7840b66e4671ba&_acct=C000059713&_version=1&_userid=108429&md5=e30c72deacbd2cd15ee7dd556b8157dd" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">Cited By in Scopus (26)</a></div><p></p><p><a name="bib5"></a></p><div id="ref_bib5"><a onclick="toggleTabs('fullTab')" href="#bbib5">5.</a> U.A. Stock and J.P. Vacanti, Tissue engineering: current state and prospects. <i>Annu Rev Med</i> <strong>52</strong> (2001), pp. 443–451. <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=3&_targetURL=http%3A%2F%2Fdx.doi.org%2F10.1146%2Fannurev.med.52.1.443&_acct=C000059713&_version=1&_userid=108429&md5=512b7758d9e6edd859a81005b7db520c" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()"><b>Full Text</b> via CrossRef</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=655&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Frecord.url%3Feid%3D2-s2.0-0035129867%26partnerID%3D10%26rel%3DR3.0.0%26md5%3Dcabca79182b61395b258bf5599112deb&_acct=C000059713&_version=1&_userid=108429&md5=22680bcd9dc2759f63291a96eb02df2a" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">View Record in Scopus</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=656&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Fcitedby.url%3Feid%3D2-s2.0-0035129867%26partnerID%3D10%26rel%3DR3.0.0%26md5%3Dcabca79182b61395b258bf5599112deb&_acct=C000059713&_version=1&_userid=108429&md5=17fffdc7690c0074bb63f058ffc30b2c" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">Cited By in Scopus (139)</a></div><p></p><p><a name="bib6"></a></p><div id="ref_bib6"><a onclick="toggleTabs('fullTab')" href="#bbib6">6.</a>
T. Maemura, M. Shin, M. Sato, H. Mochizuki and J.P. Vacanti, A
tissue-engineered stomach as a replacement of the native stomach. <i>Transplantation</i> <strong>76</strong> 1 (2003), pp. 61–65. <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=3&_targetURL=http%3A%2F%2Fdx.doi.org%2F10.1097%2F01.TP.0000068903.63554.1B&_acct=C000059713&_version=1&_userid=108429&md5=abbe429e49daf64ec82abab4b166f59c" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()"><b>Full Text</b> via CrossRef</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=655&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Frecord.url%3Feid%3D2-s2.0-0037623972%26partnerID%3D10%26rel%3DR3.0.0%26md5%3D03f9f8bb821c09f925344c3669557a73&_acct=C000059713&_version=1&_userid=108429&md5=be6fffe4d2567f512105e2b01fc32655" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">View Record in Scopus</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=656&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Fcitedby.url%3Feid%3D2-s2.0-0037623972%26partnerID%3D10%26rel%3DR3.0.0%26md5%3D03f9f8bb821c09f925344c3669557a73&_acct=C000059713&_version=1&_userid=108429&md5=8f702183a9e6387aeb0d9eacfc2783c4" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">Cited By in Scopus (8)</a></div><p></p><p><a name="bib7"></a></p><div id="ref_bib7"><a onclick="toggleTabs('fullTab')" href="#bbib7">7.</a>
T. Grikscheit, E.R. Ochoa, A. Srinivasan, H. Gaissert and J.P. Vacanti,
Tissue-engineered esophagus: experimental substitution by onlay patch
or interposition. <i>J Thorac Cardiovasc Surg</i> <strong>126</strong> 2 (2003), pp. 537–544. <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6WMF-4991FX6-2D&_user=108429&_coverDate=08%2F31%2F2003&_fmt=full&_orig=search&_cdi=6933&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=510f6e044b2a9060db8b7a0368db96a4&ref=full"><b>Article</b></a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiImageURL&_imagekey=B6WMF-4991FX6-2D-R&_cdi=6933&_user=108429&_check=y&_orig=search&_coverDate=08%2F31%2F2003&view=c&wchp=dGLbVzW-zSkWA&md5=0f4cb7ad4ddbeda67b8d4058096211d0&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (466 K)</a>
| <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=655&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Frecord.url%3Feid%3D2-s2.0-0041353730%26partnerID%3D10%26rel%3DR3.0.0%26md5%3D729f824ecc1ee7590405ef6dcdcd2ff9&_acct=C000059713&_version=1&_userid=108429&md5=92c2fc0ac57b39a0e836e00227308eff" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">View Record in Scopus</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=656&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Fcitedby.url%3Feid%3D2-s2.0-0041353730%26partnerID%3D10%26rel%3DR3.0.0%26md5%3D729f824ecc1ee7590405ef6dcdcd2ff9&_acct=C000059713&_version=1&_userid=108429&md5=e54ff1781babde7a4f0e2df116d0d7bb" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">Cited By in Scopus (25)</a></div><p></p><p><a name="bib8"></a></p><div id="ref_bib8"><a onclick="toggleTabs('fullTab')" href="#bbib8">8.</a>
M.P. Vacanti, J.L. Leonard, B. Dore, L.J. Bonassar, Y. Cao, S.J.
Stachelek, J.P. Vacanti, F. O’Connell, C.S. Yu, A.P. Farwell and C.A.
Vacanti, Tissue-engineered spinal cord. <i>Transplant P</i> <strong>33</strong> 1–2 (2001), pp. 592–598. <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6VJ0-42M793F-B1&_user=108429&_coverDate=03%2F31%2F2001&_fmt=full&_orig=search&_cdi=6080&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=d8bf29f0c2ebfe74b6524601dc6708ba&ref=full"><b>Article</b></a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiImageURL&_imagekey=B6VJ0-42M793F-B1-F&_cdi=6080&_user=108429&_check=y&_orig=search&_coverDate=03%2F31%2F2001&view=c&wchp=dGLbVzW-zSkWA&md5=7ba7b5741269597f34f38fb3c635c608&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (540 K)</a>
| <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=655&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Frecord.url%3Feid%3D2-s2.0-0035080539%26partnerID%3D10%26rel%3DR3.0.0%26md5%3Df94a2e4ec91bc56a6b55ac2ca63e9f00&_acct=C000059713&_version=1&_userid=108429&md5=1cc6e16948ffd68f56ae236c44f2dc43" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">View Record in Scopus</a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=RedirectURL&_method=outwardLink&_partnerName=656&_targetURL=http%3A%2F%2Fwww.scopus.com%2Finward%2Fcitedby.url%3Feid%3D2-s2.0-0035080539%26partnerID%3D10%26rel%3DR3.0.0%26md5%3Df94a2e4ec91bc56a6b55ac2ca63e9f00&_acct=C000059713&_version=1&_userid=108429&md5=d37d3f5ca5197a0362c733c9c19be8cd" target="outwardLink" onclick="var outwardWin; outwardWin=window.open('','outwardLink','scrollbars=yes,resizable=yes,directories=yes,toolbar=yes,menubar=yes,status=yes,location=yes,width=610,height=480,screenX=10,screenY=10'); outwardWin.focus()">Cited By in Scopus (30)</a></div><p></p><p><a name="bib9"></a></p><div id="ref_bib9"><a onclick="toggleTabs('fullTab')" href="#bbib9">9.</a> J.P. Vacanti, Tissue and organ engineering: can we build intestine and vital organs?. <i>J Gastrointest Surg</i> <strong>7</strong> 7 (2003), pp. 831–835. <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=ArticleURL&_udi=B6W8T-49V9HDY-3&_user=108429&_coverDate=11%2F30%2F2003&_fmt=full&_orig=search&_cdi=6663&view=c&_acct=C000059713&_version=1&_urlVersion=0&_userid=108429&md5=d4ad1f7da4a614f585aa67dd0d53a951&ref=full"><b>Article</b></a> | <a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science?_ob=MiamiImageURL&_imagekey=B6W8T-49V9HDY-3-9&_cdi=6663&_user=108429&_check=y&_orig=search&_coverDate=11%2F30%2F2003&view=c&wchp=dGLbVzW-zSkWA&md5=6031afbc4e030857dad42e4314d5295b&ie=/sdarticle.pdf" target="newPdfWin" onclick="var newWidth=((document.body.clientWidth*90)/100); var newHeight=document.body.clientHeight; var pdfWin; pdfWin=window.open('','newPdfWin','width='+newWidth+',height='+newHeight+',resizable=yes, left=50, top=50');pdfWin.focus()"><img name="pdf" style="vertical-align: middle;" src="science-direct_files/icon_pdf.gif" alt="" border="0"> PDF (357 K)</a>
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<a href="http://www.sciencedirect.com.ezproxy.lib.utexas.edu/science/journal/01429612"><b>Biomaterials</b></a><br>
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Volume 26, Issue 1</a>,
January 2005,
Pages 93-99
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