State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, New York, NY, USA.
Methods Mol Biol. 2020;2117:245-263. doi: 10.1007/978-1-0716-0301-7_15.
Neural stem cells (NSCs) have the features of both neural progenitors and stem cells, and show great potentials in translational research and regenerative medicine. Studies on NSCs have been greatly accelerated by the introduction of induced neural stem cells (iNSCs). The iNSCs are usually differentiated from induced pluripotent stem cells (iPSCs) or transdifferentiated from somatic cells such as fibroblasts or glial cells. Here, we describe a detailed protocol to reprogram human and mouse fibroblasts into iNSCs by overexpression of a transcription factor Ptf1a delivered by lentiviruses. The obtained iNSC lines have a strong self-renewal ability and are capable of differentiating into various types of neurons, astrocytes, and oligodendrocytes both in vitro and in vivo. The protocol is quite simple but powerful to produce iNSC lines.
神经干细胞(NSCs)具有神经祖细胞和干细胞的特征,在转化研究和再生医学中具有巨大的潜力。诱导性神经干细胞(iNSCs)的引入极大地加速了 NSCs 的研究。iNSCs 通常是由诱导多能干细胞(iPSCs)分化而来,或者由成纤维细胞或神经胶质细胞等体细胞转分化而来。在这里,我们描述了一种通过慢病毒转导转录因子 Ptf1a 来将人源和鼠源成纤维细胞重编程为 iNSCs 的详细方案。得到的 iNSC 系具有很强的自我更新能力,并能在体外和体内分化为各种类型的神经元、星形胶质细胞和少突胶质细胞。该方案非常简单,但却非常强大,可用于产生 iNSC 系。