Marro Samuele, Yang Nan
Department of Pathology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 265 Campus Drive, 3rd Floor, Stanford, CA, 94305, USA,
Methods Mol Biol. 2014;1150:237-46. doi: 10.1007/978-1-4939-0512-6_16.
Nuclear reprogramming by defined transcription factors became of broad interest in 2006 with the work of Takahashi and Yamanaka (Cell 126:663-676, 2006), but the first example of cell fate reshaping via ectopic expression of transcription factor was provided back in 1987 when Davis and colleagues induced features of a muscle cell in fibroblast using the muscle transcription factor MyoD (Davis et al., Cell 51:987-1000, 1987). In 2010 our laboratory described how forced expression of the three neuronal transcription factors Ascl1, Brn2, and Myt1l rapidly converts mouse fibroblasts into neuronal cells that exhibit biochemical and electrophysiological properties of neurons. We named these cells induced neuronal cells (iN cells) (Vierbuchen et al., Nature 463:1035-1041, 2010; Vierbuchen and Wernig, Nat Biotechnol 29:892-907, 2011). Interestingly, iN cells can also be derived from defined endodermal cells such as primary hepatocytes, suggesting the existence of a more general reprogramming paradigm (Marro et al., Cell Stem Cell 9:374-382, 2011). In this chapter we describe the detailed methods used to attain the direct conversion.
2006年，高桥和山中伸弥的研究成果（《细胞》126卷：663 - 676页，2006年）使通过特定转录因子进行核重编程引起了广泛关注，但早在1987年，当戴维斯及其同事利用肌肉转录因子MyoD在成纤维细胞中诱导出肌肉细胞特征时，就提供了通过异位表达转录因子重塑细胞命运的首个例子（戴维斯等人，《细胞》51卷：987 - 1000页，1987年）。2010年，我们实验室描述了如何通过强制表达三种神经元转录因子Ascl1、Brn2和Myt1l，迅速将小鼠成纤维细胞转化为具有神经元生化和电生理特性的神经元细胞。我们将这些细胞命名为诱导神经元细胞（iN细胞）（维布兴等人，《自然》463卷：1035 - 1041页，2010年；维布兴和韦尔尼希，《自然生物技术》29卷：892 - 907页，2011年）。有趣的是，iN细胞也可以从特定的内胚层细胞如原代肝细胞中获得，这表明存在一种更普遍的重编程模式（马罗等人，《细胞干细胞》9卷：374 - 382页，2011年）。在本章中，我们描述了用于实现直接转化的详细方法。
