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恒河猴胚胎在添加血清培养基中的植入前差异发育。

Differential preimplantation development of rhesus monkey embryos in serum-supplemented media.

作者信息

Weston A M, Wolf D P

机构信息

Division of Reproductive Sciences, Oregon Regional Primate Research Center, Beaverton 97006, USA.

出版信息

Mol Reprod Dev. 1996 May;44(1):88-92. doi: 10.1002/(SICI)1098-2795(199605)44:1<88::AID-MRD10>3.0.CO;2-X.

Abstract

Several media, some augmented with amino acids, have been formulated recently, based on simplex optimization, to support the preimplantation development of mouse embryos. For the highly limited studies on preimplantation development of nonhuman primate embryos, a complex medium (CMRL-1066) has been employed. Our objective was to compare the developmental ability of rhesus monkey embryos in a simple medium containing amino acids, KSOM/AA, with the complex media used previously. Zygotes (99) were recovered following in vitro fertilization (IVF) from six monkeys, allocated to either CMRL or KSOM/AA both containing 10% fetal calf serum (FCS), and monitored daily until reaching the expanded or hatched blastocyst stage. The distribution of cells between the inner cell mass (ICM) and trophectoderm was determined at the end of culture by differential nuclear staining. Although a greater number of embryos cultured in KSOM/AA vs. CMRL developed to the morula stage (80%) and beyond (66% to expanded blastocyst), the differences were not significant. Such embryos in KSOM/AA did, however, develop at a significantly faster rate, on average, reaching the expanded blastocyst stage 26 hr earlier than did embryos cultured in CMRL. KSOM/AA embryos hatched in less time and had a higher percentage (43 vs. 34) of cells allocated to the ICM. These results indicate that a simple medium, KSOM/AA, in the presence of serum, supports the development of rhesus monkey embryos at high efficiency and at a faster rate than that observed for embryos cultured in the complex medium, CMRL-1066.

摘要

最近,基于单纯形优化法配制了几种添加氨基酸的培养基,以支持小鼠胚胎的植入前发育。对于非人类灵长类胚胎植入前发育的研究非常有限,一直采用一种复杂培养基(CMRL - 1066)。我们的目的是比较恒河猴胚胎在含氨基酸的简单培养基KSOM/AA中与先前使用的复杂培养基的发育能力。从六只猴子体外受精(IVF)后回收了99个合子,将其分配到均含有10%胎牛血清(FCS)的CMRL或KSOM/AA中,每天进行监测,直至达到扩张囊胚或孵化囊胚阶段。培养结束时,通过核差异染色确定内细胞团(ICM)和滋养外胚层之间的细胞分布。虽然与在CMRL中培养的胚胎相比,在KSOM/AA中培养至桑椹胚阶段(80%)及之后(扩张囊胚66%)的胚胎数量更多,但差异不显著。然而,在KSOM/AA中的此类胚胎发育速度明显更快,平均而言,比在CMRL中培养的胚胎提前26小时达到扩张囊胚阶段。KSOM/AA胚胎孵化所需时间更短,分配到ICM的细胞百分比更高(43%对34%)。这些结果表明,在有血清存在的情况下,简单培养基KSOM/AA能够高效且快速地支持恒河猴胚胎的发育,其效率高于在复杂培养基CMRL - 1066中培养的胚胎。

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