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在三种输卵管上皮细胞共培养系统中使用简单培养基培养牛1-2细胞胚胎。

Bovine 1-2-cell embryo development using a simple medium in three oviduct epithelial cell coculture systems.

作者信息

Ellington J E, Carney E W, Farrell P B, Simkin M E, Foote R H

机构信息

Department of Animal Science, Cornell University, Ithaca, New York 14853-4801.

出版信息

Biol Reprod. 1990 Jul;43(1):97-104. doi: 10.1095/biolreprod43.1.97.

Abstract

These studies were designed to develop a coculture system using a simple medium to promote development of 1-cell bovine embryos through the 8-16-cell stage to morula and blastocyst stages. Monolayers for coculture were prepared from bovine oviduct epithelial cells (BOEC). In vivo-fertilized 1-2-cell embryos and ova (384) were surgically collected from superovulated cows. In Experiment 1, embryos cocultured in a simple glucose-free and serum-free medium (CZB) developed with superior scores of embryo quality than embryos cocultured in Ham's F-10 with serum, and a greater percentage developed past 8-16 cells than embryos cocultured in CMRL-1066 with serum (p less than 0.05). In Experiment 2, embryos cocultured with fresh BOEC monolayers averaged more (p less than 0.05) cells than did embryos in coculture with frozen-thawed BOEC monolayers or in BOEC-conditioned medium. Without glucose in the simple medium for the first 48 h of culture, more embryos blastulated (p less than 0.01) by Day 5.5 of culture (Day 6.5 of donor's estrous cycle) than embryos in the same medium with glucose present throughout. In Experiment 3, more embryos tended to hatch in BOEC coculture (p less than 0.10) than in conditioned medium. These results show that a chemically simple medium with fresh BOEC monolayers can provide a significant benefit for coculture of early bovine embryos.

摘要

这些研究旨在开发一种共培养系统,该系统使用简单培养基促进1细胞期牛胚胎从8-16细胞阶段发育至桑葚胚和囊胚阶段。共培养的单层细胞由牛输卵管上皮细胞(BOEC)制备。通过手术从超排母牛体内收集体内受精的1-2细胞胚胎和卵子(384个)。在实验1中,与在含血清的Ham's F-10培养基中培养的胚胎相比,在无葡萄糖和无血清的简单培养基(CZB)中培养的胚胎,其胚胎质量评分更高,并且发育至8-16细胞阶段之后的胚胎百分比高于在含血清的CMRL-1066培养基中培养的胚胎(p<0.05)。在实验2中,与用冻融BOEC单层细胞或BOEC条件培养基共培养的胚胎相比,与新鲜BOEC单层细胞共培养的胚胎平均细胞数更多(p<0.05)。在培养的前48小时,简单培养基中不含葡萄糖时,到培养第5.5天(供体发情周期第6.5天)发育成囊胚的胚胎比在整个培养过程中都含有葡萄糖的相同培养基中的胚胎更多(p<0.01)。在实验3中,与条件培养基相比,在BOEC共培养中更多胚胎倾向于孵化(p<0.10)。这些结果表明,含有新鲜BOEC单层细胞的化学成分简单的培养基可为早期牛胚胎的共培养提供显著益处。

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