Alexandrova Evguenia M, Moll Ute M
Department of Pathology, Stony Brook University, Stony Brook, NY, USA.
Methods Mol Biol. 2013;962:157-64. doi: 10.1007/978-1-62703-236-0_13.
Here we describe a method for generating induced pluripotent stem (iPS) cells from mouse embryonic fibroblasts (MEFs). Recombinant retroviruses carrying human transcription factors for Klf4, Oct3/4, Sox2, with or without c-Myc, are used to transduce early passage MEFs several times. Based on morphologic criteria, the resulting iPS colonies are picked manually at first, and then propagated and expanded by standard methods. iPS cells can then be differentiated into virtually any cell type or lineage, thus allowing for discoveries of new functions of p53 and mutant p53.
在此，我们描述一种从小鼠胚胎成纤维细胞（MEF）中生成诱导多能干细胞（iPS细胞）的方法。携带Klf4、Oct3/4、Sox2这几种人类转录因子（有或没有c-Myc）的重组逆转录病毒用于多次转导早期传代的MEF。首先根据形态学标准手动挑选得到的iPS集落，然后通过标准方法进行传代和扩增。随后，iPS细胞几乎可以分化为任何细胞类型或谱系，从而有助于发现p53和突变型p53的新功能。
