Generation and characterization of human induced pluripotent stem cells.
Generation and characterization of human induced pluripotent stem cells.
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DOI：
10.1002/9780470151808.sc04a02s9
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发表时间：
2009-06-01
影响因子：
--
通讯作者：
Yamanaka, Shinya
中科院分区：
文献类型：
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作者：
Ohnuki, Mari;Takahashi, Kazutoshi;Yamanaka, Shinya
本单元描述了如何生成人类诱导多能干细胞（iPS）并评估生成的iPS细胞的质量。人类诱导多能干细胞的建立和维持方法与小鼠诱导多能干细胞相似，但并不完全相同。此外，这些方案包括由胚胎干细胞研究人员建立的人类iPS细胞传代和低温保存的优秀程序，这使得培养人类iPS细胞变得容易。此外，我们还包括表征iPS细胞的方法，以供进一步研究。利用RT-PCR和免疫细胞化学检测多能性细胞标记物、胚状体分化和畸胎瘤分化分别确定体外和体内的多能性。
This unit describes how to generate human induced pluripotent stem (iPS) cells and evaluate the qualities of the generated iPS cells. The methods for establishment and maintenance of human iPS cells are similar to those for mouse iPS cells but not identical. In addition, these protocols include excellent procedures for passaging and cryopreservation of human iPS cells established by ES cell researchers, which result in an easy way to culture human iPS cells. Moreover, we include methods for characterizing iPS cells for further research. RT-PCR and immunocytochemistry for detection of pluripotent cell markers, embryoid body differentiation, and teratoma differentiation are used to determine pluripotency in vitro and in vivo, respectively.