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在输卵管细胞、胚胎成纤维细胞或STO细胞单层上共培养的绵羊胚胎的发育。

Development of ovine embryos co-cultured on oviductal cells, embryonic fibroblasts, or STO cell monolayers.

作者信息

Rexroad C E, Powell A M

机构信息

U.S. Department of Agriculture, Beltsville Agricultural Research Center, Maryland 20705.

出版信息

Biol Reprod. 1993 Oct;49(4):789-93. doi: 10.1095/biolreprod49.4.789.

Abstract

One- and two-cell ovine embryos were co-cultured on primary monolayer cultures of ovine oviductal cells (OM) and ovine embryonic fibroblasts (EF) or on monolayers of STO cells (STO), a permanent cell line, to determine whether a co-culture system could be developed for ovine embryos utilizing a well-characterized cell line. More than 65% (n = 64) of embryos co-cultured on OM and STO for 5 days cleaved beyond the "in vitro" block whereas only 26% (n = 35) of embryos co-cultured on EF cleaved to the same degree (p < 0.05). Mitotic inactivation of the monolayer did not alter the response to each cell type. In a second experiment, development of embryos was similar after co-culture on OM or STO cells for both 3 and 6 days. Co-culture of zygotes on OM and STO cells produced 38 and 33% blastocysts after 6 days of co-culture. After embryo transfer, only recipients receiving at least one blastocyst became pregnant. About 33% of the transferred blastocysts produced fetuses. STO cell co-culture may provide the same stimulus to development as OM cell co-culture and may be advantageous for study of the requirements for early ovine embryonic development. In addition, STO cells displayed contact inhibition and formed monolayers that did not overgrow embryos as did primary cultures of ovine oviductal cells.

摘要

将单细胞和双细胞绵羊胚胎与绵羊输卵管细胞(OM)和绵羊胚胎成纤维细胞(EF)的原代单层培养物或与一种永生化细胞系STO细胞的单层培养物共同培养,以确定是否可以利用一种特性明确的细胞系为绵羊胚胎开发一种共培养系统。在OM和STO上共同培养5天的胚胎中,超过65%(n = 64)的胚胎在“体外”阻滞期后发生了分裂,而在EF上共同培养的胚胎中只有26%(n = 35)分裂到相同程度(p < 0.05)。单层细胞的有丝分裂失活并未改变对每种细胞类型的反应。在第二个实验中,在OM或STO细胞上共同培养3天和6天后,胚胎的发育情况相似。合子在OM和STO细胞上共同培养6天后,分别有38%和33%发育成囊胚。胚胎移植后,只有接受至少一个囊胚的受体怀孕。约33%移植的囊胚发育成胎儿。STO细胞共培养对胚胎发育的刺激作用可能与OM细胞共培养相同,且可能有利于研究绵羊早期胚胎发育的需求。此外,STO细胞表现出接触抑制,形成的单层不会像绵羊输卵管细胞的原代培养那样过度生长覆盖胚胎。

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