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远交系小鼠单细胞胚胎的体外发育:培养基成分的影响

In vitro development of one-cell embryos from outbred mice: influence of culture medium composition.

作者信息

Spindle A

机构信息

Laboratory of Radiobiology and Environmental Health, University of California, San Francisco 94143.

出版信息

In Vitro Cell Dev Biol. 1990 Feb;26(2):151-6. doi: 10.1007/BF02624106.

Abstract

One-cell embryos from outbred mice (CF1, CD-1, and Dub:ICR) were cultured in various modifications of egg culture medium (ECM). The best development was observed in medium in which inorganic salts of modified T6 medium (mT6) replaced those of ECM. In this modification (TE), 66% of one-cell CF1 embryos developed into blastocysts, compared to 46 and 43% for ECM and mT6, respectively. Moreover, the cell numbers of blastocysts developing in TE (7.49 +/- 3.3) were higher than the cell numbers of those developing in ECM (55.1 +/- 2.4). The culture requirements of embryos varied between different stocks of mice: Fewer CF1 embryos developed to the blastocyst stage than either Dub:ICR embryos (90%) or CD-1 embryos (84%). Lowering the osmolarity of the medium from 300 to 280 mOSM, increasing the concentration of KC1 from 1.42 to 25 mM, or omitting lactate from the medium during Day 1 of culture did not further improve development of embryos, in contrast to previous reports. However, the time at which embryos were transferred to outgrowth medium influenced their postblastocyst development. The best development was observed when embryos were transferred on Day 4 of culture at the late morula-early blastocyst stage.

摘要

将远交系小鼠(CF1、CD-1和Dub:ICR)的单细胞胚胎在多种改良的卵培养液(ECM)中培养。在改良T6培养液(mT6)的无机盐替代了ECM的无机盐的培养液中观察到最佳发育情况。在这种改良培养液(TE)中,66%的CF1单细胞胚胎发育成囊胚,相比之下,ECM和mT6分别为46%和43%。此外,在TE中发育的囊胚细胞数(7.49±3.3)高于在ECM中发育的囊胚细胞数(55.1±2.4)。不同品系小鼠胚胎的培养需求有所不同:发育到囊胚阶段的CF1胚胎比Dub:ICR胚胎(90%)或CD-1胚胎(84%)少。与之前的报道相反,将培养液的渗透压从300降低到280 mOSM,将KCl浓度从1.42增加到25 mM,或在培养第1天从培养液中省略乳酸,并没有进一步改善胚胎的发育。然而,胚胎转移到生长培养液的时间会影响其囊胚后发育。当胚胎在培养第4天处于晚期桑椹胚-早期囊胚阶段转移时,观察到最佳发育情况。

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