Generation of human-induced pluripotent stem cells
Generation of human-induced pluripotent stem cells
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DOI：
10.1038/nprot.2008.92
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Daley, George Q.
中科院分区：
文献类型：
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作者：
Park, In-Hyun;Lerou, Paul H.;Daley, George Q.
多能细胞，如胚胎干细胞，是研究的宝贵工具，并有可能作为细胞和组织替代疗法的来源。胚胎干细胞来源的细胞和组织移植后的排斥反应是其临床应用的重大障碍。最近，通过异位表达四种转录因子（Oct4、Sox2、Klf4和Myc）将人类体细胞直接重编程为多能性，以产生诱导的多能干（iPS）细胞。人类iPS细胞是患者特异性多能干细胞的潜在来源，可以绕过免疫排斥反应。iPS细胞也可用于研究没有适当的人类体外或动物模型的疾病。在本方案中，我们描述了如何建立原代人成纤维细胞系以及如何通过逆转录病毒转导重编程因子来获得iPS细胞。总体而言，从活检开始，完成人类原代成纤维细胞重编程需要2个月的时间。
Pluripotent cells, such as embryonic stem cells, are invaluable tools for research and can potentially serve as a source of cell- and tissue-replacement therapy. Rejection after transplantation of cells and tissue derived from embryonic stem cells is a significant obstacle to their clinical use. Recently, human somatic cells have been reprogrammed directly to pluripotency by ectopic expression of four transcription factors (Oct4, Sox2, Klf4 and Myc) to yield induced pluripotent stem (iPS) cells. Human iPS cells are a potential source of patient-specific pluripotent stem cells that would bypass immune rejection. iPS cells can also be used to study diseases for which there are no adequate human in vitro or animal models. In this protocol, we describe how to establish primary human fibroblasts lines and how to derive iPS cells by retroviral transduction of reprogramming factors. Overall, it takes 2 months to complete reprogramming human primary fibroblasts starting from biopsy.