Procedures for In Vitro Production of Bovine Embryos

R.M Rivera, A.M. Brad, J.L. Edwards, A.D. Ealy, V.M. Monterroso,
A.C. Majewski, C.M. Franco, J.M. Moss, J.M. Fear, and P.J. Hansen
Dept. of Animal Sciences, University of Florida

The procedures for in vitro production [IVP; i.e. in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC)] of embryos described here are based on procedures developed in other laboratories at the University of Wisconsin (Parrish et al., 1986), University of Guelph  (Xu et al., 1992), and University of Missouri (Hernandez-Ledezma et al., 1993).  These procedures as used by our group have been published previously (Edwards et al., 1997; Paula-Lopes et al., 1998).  Keep in mind that the protocols described here are not fixed but rather constantly evolve as new developments take place.  Therefore, practitioners of IVP will be well advised to experiment with the procedures used, especially after reading of improvements made by other laboratories.  This protocol is organized by day of the protocol with d 0 being the day of fertilization.

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DAY-BY-DAY PROCEDURE
Day -2:  Getting Prepared
Day -1:  Collection of Ovaries | Oocyte Collection | Preparation of IVF Medium
Day 0:   In Vitro Fertilization
Day 1-9:  Culture of Embryos

Abbreviated Protocol for the Experienced User
Instructions for Printing This Manual        Obtaining The Manual in PDF Format

Abbreviated Protocol in Spanish (thanks Adriana Fernandez)

Detailed Protocol in Spanish (protocol by Luis Luis Armando Dávila Félix - thanks Luis)

Abbreviated Protocol in Portuguese (thanks Rodrigo Nunes and Luiz de Castro e Paula)

OTHER INFORMATION

Click on the icon to see recent changes in the IVP Protocol

Do you want a hard copy of our embryo poster? Drop a line by email to Peter Hansen or Sabrina Robinson with your shipping address and we will send you at no charge an 8.5" x 11" color poster illustrating all of the developmental endpoints observed during IVP of bovine embryos. 

Contact the Author:    Hansen.  We want your feedback!! Let us know whether this protocol works for you.

The image shown on this page is of a day 6 bovine embryo subjected to staining using the TUNEL reagent and propidium iodide and analyzed using a Zeiss Axioplan microscope using DIC. The nucleus staining yellow is positive for apoptosis. Image by Paolete Soto (© Soto and Hansen, 2002).

This page is a contribution to the International Dairy Heat Stress Consortium and was funded in part by a grant from the USDA Initiative for Future Agricultural and Food Systems (USDA CSREES# 2001-52101-11318).

All original text and images in this site © Rivera, Hansen, et al., 2000-2009
 this site was created 4-10-00.  This page was last updated  3-31-09

Links to commercial sites do not constitute endorsement by the authors or the University of Florida