Vasil'ev I M, Vasil'eva S G
Ontogenez. 1995 May-Jun;26(3):201-5.
To study factors influencing the preparation of porcine ES cells, intact pig embryos at various stages of preimplantation development, as well as clusters of embryonic cells prepared from embryos at the stage of late or expanded blastocyst were cultivated in alpha-MEM, alpha-MEM + 50% BRL-conditioned medium, or in PBS + 20% FCS. NIH 3T3 cells, or pig granulosa cells were used as a feeder layer. It has been demonstrated that incubation conditions seriously affect development, release, and attachment to the feeder layer of both intact pig embryos and clusters of embryonic cells. However, the stage of preimplantation development is the most important factor, defining success in preparation of pig ES cells. It has been shown that embryoblast of pig embryos at stages of early, middle, late, and expanded blastocyst consists of 4-5, 8-10, 12-15, and 16-18 cells, respectively. This leads us to the conclusion that the rate of proliferation of ICM cells from pig embryos is very low. These results indirectly confirm the existence in embryonic development of pigs of metabolically resting ICM cells up to the beginning of gastrulation.
为研究影响猪胚胎干细胞制备的因素，将处于不同着床前发育阶段的完整猪胚胎以及由晚期或扩张囊胚阶段的胚胎制备的胚胎细胞团，分别在α - MEM、α - MEM + 50% BRL条件培养基或PBS + 20% FCS中培养。使用NIH 3T3细胞或猪颗粒细胞作为饲养层。结果表明，培养条件严重影响完整猪胚胎和胚胎细胞团向饲养层的发育、释放及附着。然而，着床前发育阶段是决定猪胚胎干细胞制备成功与否的最重要因素。研究显示，早期、中期、晚期和扩张囊胚阶段猪胚胎的内细胞团分别由4 - 5个、8 - 10个、12 - 15个和16 - 18个细胞组成。由此我们得出结论，猪胚胎内细胞团细胞的增殖速率非常低。这些结果间接证实了在猪胚胎发育过程中，直到原肠胚形成开始前，存在代谢静止的内细胞团细胞。
