重编程肝癌细胞系Huh7为多潜能干细胞样细胞

[摘要] 目的 重编程肝癌细胞系Huh7细胞为多潜能干细胞。方法 包装携带有Oct4、Sox2、Nanog、Lin28基因的慢病毒，将包装好的病毒共感染Huh7细胞，采用碱性磷酸酶染色、免疫荧光、实时定量PCR，免疫组织化学等方法对诱导出的多潜能干细胞样克隆细胞进行鉴定。结果 Huh7细胞被重编程为多潜能干细胞样细胞，碱性磷酸酶染色呈阳性，免疫荧光实验证明其表达多潜能因子，Oct4与TRA-1-60，qPCR实验证明其高水平表达内源性的多潜能相关基因及干细胞特异的microRNAs，体内分化实验结果表明iHuh7可以形成畸胎瘤。结论 通过携带四种多潜能因子的慢病毒介导的重编程， Huh7细胞可以被诱导为多潜能干细胞样细胞。

Reprogramming hepatocellular carcinoma cell Huh7 into pluripotent stem-like cells

[Abstract] Objective To reprogram the human hepatocellular carcinoma cell Huh7 into pluripotent stem cells. Methods Four recombinant lentivirus individually carrying Oct4, Sox2, Nanog, and Lin28 were constructed and used to co-infect Huh7 cells in vitro. Post infection, the obtained pluripotent stem-like cells were identified by Alkaline phosphatase staining, immunofluorescence assay, quantitative-PCR and immunohistochemistry . Results After lentivirus-based induction, pluripotent stem-like cell colonies could be observed in cultured Huh7 cells. These colonies showed positive in alkaline phosphatase staining and were expressing the pluripotent factors, Oct4 and TRA-1-60. Quantitative-PCR indicated that several endogenous pluripotency-associated genes and stem cell-specific microRNAs were highly expressing in these pluripotent stem-like cells. Conclusions Huh7 cells could be induced into pluripotent stem-like cells,which are mediated by lentivirus individually carrying Oct4, Sox2, Nanog, and Lin28.