Krivokharchenko A S, Vil'ianovich L I, Tatarinova L V, Riabykh V P
Ontogenez. 1993 Nov-Dec;24(6):53-60.
We studied the development of mouse embryos in vitro depending on the number of embryos in a given microvolume of the Ham's F-10 medium without protein or with the addition of serum. The absence of serum from the culture medium did not affect the development of two-cell embryos cultivate in groups of 5-6 (about 90% embryos developed until the stage of blastocyst and over 50% left zona pellucida), but the development of single embryos in the protein-free medium proceeded significantly worse. Single two-cell embryos cultivated in the serum-containing medium developed similar to embryos cultivated in groups. At the same time, no significant differences in development was found between eight-cell embryos cultivated individually or in groups of up to 10 embryos in the Ham's F-10 medium, either in the presence of serum or without it (about 95% embryos developed until the stage of blastocyst and over 70% left zona pellucida). The increase in the number of cultivated embryos over 10 had the adverse effect on development of either two-cell or eight-cell embryos. The attachment of blastocysts to the substrate after leaving zona pellucida and growth of trophectoderm was observed only in the presence of serum. These results suggest that interaction between preimplantation embryos in culture can probably be mediated by factor(s) released by embryos into the medium. Serum appears to contain such factor(s). Sensitivity of embryos to the factor(s) apparently depends on the stage of development.
我们研究了小鼠胚胎在体外的发育情况，具体取决于在不含蛋白质或添加了血清的特定微体积Ham's F-10培养基中的胚胎数量。培养基中无血清并不影响以5至6个为一组培养的二细胞胚胎的发育（约90%的胚胎发育至囊胚阶段，超过50%的胚胎孵化），但在无蛋白质培养基中单胚胎的发育明显较差。在含血清培养基中培养的单个二细胞胚胎的发育情况与成组培养的胚胎相似。同时，在Ham's F-10培养基中，无论有无血清，单个培养或多达10个一组培养的八细胞胚胎在发育上没有显著差异（约95%的胚胎发育至囊胚阶段，超过70%的胚胎孵化）。培养胚胎数量超过10个会对二细胞或八细胞胚胎的发育产生不利影响。只有在有血清的情况下，才能观察到囊胚在孵化后附着于底物以及滋养外胚层的生长。这些结果表明，培养中植入前胚胎之间的相互作用可能由胚胎释放到培养基中的因子介导。血清似乎含有此类因子。胚胎对该因子的敏感性显然取决于发育阶段。
