Nowshari M A, Holtz W
Institute of Animal Husbandry and Genetics, University of Goettingen, FRG.
J Anim Sci. 1993 Dec;71(12):3403-8. doi: 10.2527/1993.71123403x.
Embryos in the morula to blastocyst stage were collected from superovulated goats either surgically or after slaughter. Embryos with good or fair morphology were bisected with a microblade without the aid of a holding pipette or other microinstruments. Of 103 morulae and 77 blastocysts that were split, 200 (97%) and 151 (98%) demi-embryos, respectively, with no major morphological aberrations were obtained. Zona-free demi-embryos derived from blastocysts were incubated in vitro for 2 h, and those derived from morulae for 24 h so that they would reach the blastocyst stage. Demi-embryos were transferred either fresh or after freezing and thawing. After 2 h of incubation, a significantly higher proportion of zona-free demi-embryos derived from blastocysts were of good quality than were zona-free demi- embryos derived from morulae (75 vs 45%). Seventy-six percent of zona-free demi-embryos derived from morulae developed to the blastocyst stage after 24 h in vitro. The quality of the embryos before splitting had a significant effect on the development of the demi-embryos in vitro. Transfer of 11 pairs of zona-free demi-embryos resulted in the birth of five twin and three singleton kids. Transfer of 11 pairs of demi-embryos frozen-thawed without zonae pellucidae resulted in the delivery of two singleton kids.
通过手术或屠宰后从超数排卵的山羊中收集桑葚胚至囊胚阶段的胚胎。在不借助固定吸管或其他显微器械的情况下，用微型刀片将形态良好或尚可的胚胎一分为二。在分割的103个桑葚胚和77个囊胚中，分别获得了200个（97%）和151个（98%）无明显形态异常的半胚。将来自囊胚的去透明带半胚体外培养2小时，来自桑葚胚的半胚体外培养24小时，使其发育至囊胚阶段。半胚可新鲜移植，也可冷冻解冻后移植。培养2小时后，来自囊胚的去透明带半胚质量良好的比例显著高于来自桑葚胚的去透明带半胚（75%对45%）。来自桑葚胚的去透明带半胚在体外培养24小时后，76%发育至囊胚阶段。分割前胚胎的质量对半胚的体外发育有显著影响。移植11对去透明带半胚，产下了5对双胞胎和3只单胎羔羊。移植11对去透明带冷冻解冻后的半胚，产下了2只单胎羔羊。
