Transient folate deprivation in combination with small-molecule compounds facilitates the generation of somatic cell-derived pluripotent stem cells in mice
Transient folate deprivation in combination with small-molecule compounds facilitates the generation of somatic cell-derived pluripotent stem cells in mice
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DOI：
10.1007/s11596-014-1249-5
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发表时间：
2014-04
期刊：
影响因子：
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通讯作者：
Wen-tao Hu;Qiuyue Yan;Yu Fang;Zhan-dong Qiu;Su-ming Zhang
中科院分区：
文献类型：
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作者：
Wen-tao Hu;Qiuyue Yan;Yu Fang;Zhan-dong Qiu;Su-ming Zhang
诱导多能干细胞（iPSC）可以无限繁殖，同时保持分化为除胚胎外组织外的体内所有细胞类型的能力。这种iPSC技术不仅代表了一种将个体特异性干细胞用于再生医学的新方法，而且还构成了一种获得大量疾病特异性细胞用于生物医学研究的新方法。然而，低效率的重编程和癌基因和病毒载体的基因组整合限制了iPSCs的潜在应用。化学诱导的重编程提供了一种产生iPSC的新方法。在这项研究中，在瞬时叶酸剥夺条件下，使用小分子化合物（SM）（丁酸钠，A-83-01，CHIR 99021，Y-27632）的新组合来产生iPSC。发现瞬时叶酸剥夺与SM组合足以允许在转录因子Oct 4和Klf 4存在下在25天内从小鼠胚胎成纤维细胞（MEF）重编程，取代Sox 2和c-Myc，并加速小鼠iPSC的产生。所得细胞系在增殖速率、形态、多能性相关标志物和基因表达方面类似于小鼠胚胎干（ES）细胞。叶酸的去除，结合用SM处理MEFs，可以提高iPSC的诱导效率，并减少其致癌性和外源性重编程因子的使用。
Induced pluripotent stem cells (iPSCs) can be propagated indefinitely, while maintaining the capacity to differentiate into all cell types in the body except for the extra-embryonic tissues. This iPSC technology not only represents a new way to use individual-specific stem cells for regenerative medicine but also constitutes a novel method to obtain large numbers of disease-specific cells for biomedical research. However, the low efficiency of reprogramming and genomic integration of oncogenes and viral vectors limit the potential application of iPSCs. Chemical-induced reprogramming offers a novel approach to generating iPSCs. In this study, a new combination of small-molecule compounds (SMs) (sodium butyrate, A-83-01, CHIR99021, Y-27632) under conditions of transient folate deprivation was used to generate iPSC. It was found that transient folate deprivation combined with SMs was sufficient to permit reprogramming from mouse embryonic fibroblasts (MEFs) in the presence of transcription factors, Oct4 and Klf4, within 25 days, replacing Sox2 and c-Myc, and accelerated the generation of mouse iPSCs. The resulting cell lines resembled mouse embryonic stem (ES) cells with respect to proliferation rate, morphology, pluripotency-associated markers and gene expressions. Deprivation of folic acid, combined with treating MEFs with SMs, can improve the inducing efficiency of iPSCs and reduce their carcinogenicity and the use of exogenous reprogramming factors.