实验方法> 细胞技术> 细胞功能测定>An Improved Method for Generating and Identifying Human Induced Pluripotent Stem Cells

An Improved Method for Generating and Identifying Human Induced Pluripotent Stem Cells

关键词: pluripotent stem cells来源: 互联网

This chapter describes the methods we use to derive and characterize human induced pluripotent stem (iPS) cells. We describe in order, first our culture techniques for the starting fibroblast populations and methods for retrovirus preparation and concentration. Subsequently, a detailed iPS derivation protocol suitable for human fibroblast populations is discussed using standard retroviral vectors expressing the classic four or three reprogramming genes. Finally, we elaborate a robust technique for monitoring and identification of potential iPS cells through live staining of reprogrammed cells. We also outline steps for characterization of the resulting iPS cell lines.

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