Cryopreservation of Goat Embryos through Vitrification

News source: Philippine Council for Agriculture,
Forestry and Natural Resources Research and Development
For further information, see Ocampo, M. Protocol for embryo transfer and cryopreservation in goat. Los Baños, Laguna: PCARRD/DOST, 2002., 2003-03-01

Simple and Inexpensive Procedure

Cryopreservation of goat embryos used to be tedious and complicated for field use. Researchers developed a simple and inexpensive cryoperservation procedure, which enhances the feasibility of on-farm embryo cryopreservation.

Minimum Drop Size Technique

The technique, a modified vitrification procedure, is called minimum drop size technique. It involves the following steps: 1) Select morula to blastocyst stage embryos that are morphologically good to excellent in quality. 2) Wash embryos three times and maintain in a fresh holding medium. 3) Equilibrate the embryos by the two-step procedure at room temperature. 4) Place the embryos in a Petri dish (35 x 10 mm) with 2 ml vitrification solution A [VsA = 10% ethylene glycol (EG) in 20% FBS Hepes-buffered TCM-199 medium] for 10 minutes. 5) Transfer to vitrification solution B (VsB = 40% EG + 1M sucrose in 20% FBS Hepes-buffered TCM-199 medium) for 45 seconds. 6) Using a micropipette, transfer the embryos from the VsB solution into the LN2 in a styrofoam box. 7) Freeze the VsB solution with the embryos (1-2 embryos per drop) in "pellet form" (3 mm in diameter). 8) Using a thumb forceps, place the frozen embryos in a cryotube before storing in an LN2 tank for future use.

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