研究鼠胚成纤维细胞系STO (SIM-6-thiogunanie-oualiain)诱导为多功能干细胞(induced pluripotent stem cells),(STO-iPSCs).通过磷酸钙法将连接有Oct4、Sox2、Klf4和c-Myc 4个转录因子的慢病毒载体FUW-OSKM转染293FT细胞包装病毒,然后用病毒感染STO细胞,以干细胞培养条件培养.挑取诱导15d的克隆并在有饲养层和无饲养层的培养体系中培养.对获取的STO-iPSCs进行生物学特性分析,具有典型胚胎干细胞的形态特征,碱性磷酸酶染色呈阳性；qPCR结果表明,表达很高的原癌基因Nanog和Oct4 mRNAs;免疫细胞化学表明,表达胚胎干细胞特异性标志Nanog和Oct4,并且能够体外诱导分化为神经细胞.实验获得了STO-iPSCs,建立了STO-iPSCs无饲养层培养体系.%Research of the induced pluripotent stem cells (iPSCs) by inducing the SIM-6-thiogunanie-oualiain (STO) cells. Lentivirus vector FUW-OSKM which included octamer-binding transcription factor 4(Oct4), SRY (sex determining region Y)-box 2(Sox2), Kruppel-like factor 4 (Klf4) and c-Myc transfected to 293FT cells by calcium phosphate method. Then, using the virus infected STO cells after lentivirus was packaged and the cells were cultured in the ESC media. The clones were isolated on the fifteenth day cultured on feed layer and none feed layer culture dishs.Through the biological characteristics analysis, we found the cells named STO-iPSCs had the typical morphological characteristics of embryonic stem cells, had a positive result by alkaline phosphatase staining,expressed embryonic stem cells marker Nanog and Oct4 by the immunocytochemistry and high quantitys of Nanog and Oct4 mRNAs by qPCR, especially could differentiate to nerve cells in vitro. In conclution, we obtained STO-iPSCs and stablished the STO-iPSCs training system without raising layer.
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