Fukui Y
Dept. of Meat Anim. Reprod., Obihiro University of Agriculture and Veterinary Medicine, Japan.
J Anim Sci. 1989 May;67(5):1318-23. doi: 10.2527/jas1989.6751318x.
The present experiment was designed to identify possible effects of sera and steroid hormones added to a co-culture with bovine oviduct epithelial cells on embryonic development in vitro. Bovine oocytes were matured in vitro for 24 h and then fertilized in vitro using swim-up and heparin-treated, frozen-thawed spermatozoa. At 18 and 20 h after insemination, oocytes were cultured for 3 or 7 d in a co-culture system with bovine oviduct epithelial cells containing either fetal calf serum (FCS) or estrous cow serum (ECS) and one of six hormonal additions (none, 1 or 10 micrograms/ml estradiol [E]; 1 microgram/ml progesterone [P]; 1 microgram/ml E + P; and 10 micrograms/ml E + P). A total of 2,666 oocytes were cultured for 3 d and examined for cleavage. Of those, 2,280 oocytes were cultured up to 7 d for development to the late morula or blastocyst stage. Greatest cleavage rates for 2- to 8-cell and 8-cell stages were observed in FCS (71 and 24%) and ECS (66 and 23%) without steroid addition. For development into blastocysts, no serum effect was observed. Greatest rates for development into blastocysts were observed in FCS (14%) and ECS (16%) without steroid addition. These results indicate that addition of E and P at the doses and combinations tested did not enhance developmental capacity of in vitro fertilized bovine oocytes. Compared with FCS, ECS tended to increase cleavage rates and development into blastocysts.
本实验旨在确定添加到与牛输卵管上皮细胞共培养体系中的血清和类固醇激素对体外胚胎发育可能产生的影响。牛卵母细胞在体外成熟24小时,然后使用上浮法和肝素处理的冻融精子进行体外受精。授精后18和20小时,将卵母细胞在与含有胎牛血清(FCS)或发情母牛血清(ECS)的牛输卵管上皮细胞的共培养体系中培养3或7天,并添加六种激素之一(无添加、1或10微克/毫升雌二醇[E];1微克/毫升孕酮[P];1微克/毫升E + P;以及10微克/毫升E + P)。总共2666个卵母细胞培养3天并检查其分裂情况。其中,2280个卵母细胞培养至7天,观察其发育至桑葚胚晚期或囊胚期。在不添加类固醇的FCS(71%和24%)和ECS(66%和23%)中观察到2 - 8细胞期和8细胞期的最高分裂率。对于发育成囊胚,未观察到血清效应。在不添加类固醇的FCS(14%)和ECS(16%)中观察到发育成囊胚的最高比率。这些结果表明,在所测试的剂量和组合下添加E和P并未提高体外受精牛卵母细胞的发育能力。与FCS相比,ECS倾向于提高分裂率和发育成囊胚的比率。