Summary: iPS cells or induced pluripotent stem cells are stem cell-like cells derived from reprogrammed somatic cells. iPS cells reprogramming is achieved by simultaneously expressing 2-4 ES cell specific transcriptional factors including Oct4, Sox2, Myc and Klf4/Lin28 using viral vectors (lentivirus or retrovirus). The reprogramming process takes 15-30 days. First, somatic cells such as fibroblast cells are infected with a mixture of viral particles expressing the 2-4 stem cell factors. The infected cells are then seeded on mouse embryonic fibroblast (MEF) feeder cells and cultured in ES cell medium. After 10-15 days, ES like colonies of cells will appear, can be picked and expanded. Reprogramming can also be done without using MEF feeder cells.
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Protocols
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Basic iPS MEF-Based cell culture
(WiCell Research Institute)
This booklet contains MEF-based iPS cell culture protocols.
https://www.wicell.org/index.php?option=com_docman...
Added: Wed Jan 28 2009, Hits: 186, Reviews: 0 Write review -
DNA Extraction for Stem Cells
(WiCell Research Institute)
https://www.wicell.org/index.php?option=com_docman...
Added: Fri Feb 20 2009, Hits: 88, Reviews: 0 Write review -
Induced Pluripotent Stem (iPS) Cell Culture
(WiCell Research Institute)
Protocols for culturing established iPS cells without using feeder cells.
https://www.wicell.org/index.php?option=com_docman...
Added: Fri Feb 20 2009, Hits: 158, Reviews: 0 Write review -
iPS Cell Media & Reagents
(Thomson Lab, University of Wisconsin)
http://ink.primate.wisc.edu/~thomson/protocol.html...
Added: Wed Jan 28 2009, Hits: 138, Reviews: 0 Write review Cached -
Reprogramming somatic cells using cell extracts
(Collas Lab, University of Oslo, Institute of Basic Medical Sciences)
An in vitro approach for directly turning one somatic cell type into another. The system relies on the transient uptake of regulatory components from a nuclear and cytoplasmic extract derived from a “target cell” by the nucleus of a reversibly permeabilized cell (referred to as the “donor cell”). Reprogramming extracts are prepared by sonication of a target cell type and sedimentation of the coarse material.
http://www.collaslab.com/UserFiles/File/Nuclear%20...
Added: Wed Jan 28 2009, Hits: 194, Reviews: 0 Write review -
Splitting iPS cells on MEFs
(Thomson Lab, University of Wisconsin)
http://ink.primate.wisc.edu/~thomson/protocol.html...
Added: Wed Jan 28 2009, Hits: 127, Reviews: 0 Write review Cached