Filia F, Loi P, Ledda S, Branca A, Fadda P
Istituto di Fisiologia Veterinaria, Università di Sassari.
Boll Soc Ital Biol Sper. 1990 Dec;66(12):1165-71.
Embryos obtained from Sardinian breed ewes superovulated with FSH-p (Sigma) were frozen at -196 degrees C in liquid nitrogen. After 6 months storage, the embryos (12, all at the compact morula stage), were thawed in a water bath at 39 degrees C for 30 minutes. Six embryos were dissected with a Leitz micromanipulator using a simplified technique. Both demi and intact embryos, were cultured in medium TCM 199 + 10% FCS at 38 degrees C in 5% CO2 for 24 hours. Only five demi-embryos (41.6%) became blastocysts versus 4 whole embryos (66.6%) after the culture period. Splitting as our results show, lowers the embryo viability after freezing-thawing, but it can be used in certain instances to obtain genetic improvement in the Sardinian breed.
从用FSH-p（西格玛）进行超数排卵的撒丁岛品种母羊获得的胚胎，在液氮中于-196摄氏度下冷冻。储存6个月后，将胚胎（共12个，均处于致密桑椹胚阶段）在39摄氏度的水浴中解冻30分钟。使用简化技术，用徕卡显微操作仪解剖6个胚胎。半胚和完整胚胎均在含10%胎牛血清的TCM 199培养基中，于38摄氏度、5%二氧化碳条件下培养24小时。培养期后，只有5个半胚（41.6%）发育成囊胚，而完整胚胎有4个（66.6%）发育成囊胚。如我们的结果所示，分割会降低胚胎冻融后的活力，但在某些情况下可用于撒丁岛品种的遗传改良。
