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Characterization of embryonic stem-like cells derived from HEK293T cells through miR302/367 expression and their potentiality to differentiate into germ-like cells

机译:通过miR302 / 367表达表征源自HEK293T细胞的胚胎干样细胞及其分化为生殖样细胞的潜力

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摘要

Human induced pluripotent stem (iPS) cells have great value for regenerative medicine, but are facing problems of low efficiency. MicroRNAs are a recently discovered class of 19–25 nt small RNAs that negatively target mRNAs. miR302/367 cluster has been demonstrated to reprogram mouse and human somatic cells to iPS cells without exogenous transcription factors, however, the repetition and differentiation potentiality of miR302/367-induced pluripotent stem (mirPS) cells need to be improved. Here, we showed overexpression of miR302/367 cluster reprogrammed human embryonic kidney 293T cells into mirPS cells in serum-free N2B27-based medium. The mirPS cells had similar morphology with embryonic stem cells, and expressed pluripotent markers including Oct4, Sox2, Klf4, and Nanog. In addition, through formation of embryoid bodies, various cells and tissues from three germ layers could be determined. Moreover, we examined the potential of mirPS cells differentiating into germ cells both in vitro and in vivo. Taken together, these data might provide a new source of cells and technique for the investigation of the mechanisms underlying reprogramming and pluripotency.
机译:人诱导的多能干(iPS)细胞在再生医学上具有巨大价值,但面临效率低下的问题。 MicroRNA是一类新近发现的19-25 nt的小RNA,它们负向靶向mRNA。已证明miR302 / 367簇可将小鼠和人类体细胞重编程为无外源转录因子的iPS细胞,但是,miR302 / 367诱导的多能干(mirPS)细胞的重复和分化潜能有待提高。在这里,我们显示了在无血清基于N2B27的培养基中将miR302 / 367簇重编程的人类胚胎肾脏293T细胞过度表达为mirPS细胞。 mirPS细胞的形态与胚胎干细胞相似,并表达多能性标志物,包括Oct4,Sox2,Klf4和Nanog。另外,通过形成胚状体,可以确定来自三个胚层的各种细胞和组织。此外,我们在体外和体内检查了mirPS细胞分化为生殖细胞的潜力。综上所述,这些数据可能为研究重编程和多能性的机制提供了新的细胞和技术来源。

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