小鼠ES细胞在不同饲养层上培养nanog基因的表达水平与生长行为的研究

观察小鼠ES-D3细胞在MEF、新生牛睾丸支持细胞(nBTSCs)和新生兔睾丸支持细胞(nRTSCs)饲养层上生长4dnanog基因的表达水平和生长行为。RT-PCR分析显示,MEF饲养层上培养4d的ES-D3nanog基因含量高;nBTSC饲养层上培养nanog基因表达较低;而RTSC饲养层上培养nanog基因表达最低。结果显示:不同饲养层上的ES-D3细胞集落形态和培养4d分化程度有差异。在MEF饲养层上的ES-D3细胞培养4d,集落形态仍然很规则,细胞间紧密,集落表面少见大细胞,AKP染色强阳性。在nBTSC饲养层上的ES-D3细胞,集落界限清晰,AKP染色显示,集落大部分细胞呈阳性,少数的细胞集落为弱阳性,可见集落表面分散有较多的大细胞。在nRTSC饲养层上的饲养层上的ES-D3,集落隆起不明显,集落界限不清晰,大部分集落形态不太规则,AKP染色显示集落中部分细胞呈阳性,少数的细胞集落为全阴性,集落表面有较少的大细胞,集落周围有伸出的成纤维细胞。

Nanog Expression and Growth Behavior of Mouse ES Cells on Different Feeders

It is Observed that the nanog expression level and the growth behavior of the mouse ES-D3 cells cultured on the MEF feeder, the neonatal bovine testicular sertoli cells (nBTSCs) feeder and the neonatal rabbit testicular sertoli cells (nRTSCs) feeder on the 4th day. RT-PCR shows that nanog expresses high level on the MEF feeder and lower on the nBTSCs and the nRTSCs the lowest. Results show that the colony form and the differentiation degree of the ES-D3 cells cultured on the different feeders are different. Colonies of the ES-D3 cultured on the MEF feeder have very regular form and show strong positive for AKP staining. The cells are compact. There are few big cells on the surface of the colonies. Cells on the nBTSCs feeder have clear bourn but spread around some big cells on the colonies surface. Most colonies show positive and a few show weak positive for AKP staining. Cells on the nRTSCs feeder have irregular and unclear colonies. Only part of the colonies shows positive for AKP staining and some show negative. There are a few big cells on the surface of the colonies and the fibroblast spreads around them.