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目的:通过慢病毒转染技术成功建立了人诱导分化多能干细胞系(induced pluripotent stem cell,iPSC)并诱导它们向神经上皮细胞(neuroepithelial cells,NECs)定向分化,为研究人胚胎早期神经发育建立一种新的模型,并有望为临床神经系统病变的治疗提供一种新的移植选择。方法:采用慢病毒将转录因子八聚体结合转录因子-4(octamer-binding transcription factor-4,OCT4)、性别决定Y区域转录因子2[sex determining region Y(SRY)-box 2,SOX2]、Nanog同源盒(Nanog homebox,NANOG)、Lin-28同源因子(Lin-28 homolog,LIN28)、原癌基因c-Myc和Kruppel样因子4(Kruppel-like factor 4,KLF4)导入到人胎肺成纤维细胞中,经过筛选得到重编程后的人iPSC,并在体外通过加入数种生长因子将其诱导成为NECs。结果:成功获得与成纤维细胞相同遗传背景的iPSC,具有与人胚干细胞(human embryonic stem cell,hESC)相似的形态学及多向分化潜能,且能在体外被诱导分化成为NECs,分化效率与hESC相似。结论:通过重编程技术成功建立人iPSC并将其定向诱导分化为NECs,为iPSC在神经系统疾病特别是遗传性或退行性神经病变的治疗和机制研究中的应用推广奠定了实验基础。
OBJECTIVE: To successfully establish induced pluripotent stem cells (iPSCs) by lentiviral transfection and to induce them to differentiate into neuroepithelial cells (NECs). To study the early neural development of human embryos A new model is expected to provide a new choice of transplantation for the treatment of clinical nervous system diseases. Methods: Lentivirus was transfected into octamer-binding transcription factor-4 (OCT4) and sex determining region Y (SRY) -box 2, SOX2] Nanog home box (NANOG), Lin-28 homolog (LIN28), proto-oncogene c-Myc and Kruppel-like factor 4 (KLF4) Pulmonary fibroblasts were reprogrammed to obtain reprogrammed human iPSCs and in vitro induced to become NECs by addition of several growth factors. Results: iPSCs, which had the same genetic background as fibroblasts, had the same morphological and multidirectional differentiation potential as human embryonic stem cells (hESC), and could be induced to differentiate into NECs in vitro. The differentiation efficiency Similar to hESC. CONCLUSION: Successful establishment of human iPSC through reprogramming technology and its directional differentiation into NECs lay the foundation for the promotion and application of iPSC in the treatment and mechanism research of neurological diseases, especially hereditary or degenerative neuropathy.