Ding Yi, Chang Cuifang, Niu Zhipeng, Dai Keqiang, Geng Xiaofang, Li Deming, Guo Jianlin, Xu Cunshuan
College of Life Science, Henan Normal University, Xinxiang, 453007, Henan Province, China.
State Key Laboratory Cultivation Base for Cell Differentiation Regulation and Henan Bioengineering Key Laboratory, Henan Normal University, Xinxiang, 453007, Henan Province, China.
Cytotechnology. 2016 Oct;68(5):2037-47. doi: 10.1007/s10616-016-9944-7. Epub 2016 Jan 21.
Hepatocytes differentiated from induced pluripotent stem cells and adult stem cells could be utilized as a tool for the study of liver diseases, screening for drug metabolism and hepatotoxicity. Thus further investigation of the method to efficiently generate hepatocytes is in great need. Bone Mesenchymal Stem Cells (BMSCs) were collected from rat femurs and tibias. FOXA2 and HNF1α genes were constructed into a lentiviral vector and introduced into BMSCs by a lentivirus-mediated overexpression system. Three weeks after the induction, the expressions of FOXA2 and HNF1α, and liver specific genes were analyzed, and hepatocyte-function related assays were performed. Overexpression of both FOXA2 and HNF1α induced the BMSCs to differentiate into hepatocyte-like cells (HLCs). Hepatocyte-specific gene and protein were detected by RT-PCR, Western Blot and Immunofluorescence. These HLCs also exerted some typical hepatocyte functions such as glycogen storage, indocyanine green absorption and lipid accumulation. The combination of FOXA2 and HNF1α can effectively induce BMSCs to differentiate into HLCs. This is a novel and efficient method to prepare HLCs within a short timeline.
从诱导多能干细胞和成体干细胞分化而来的肝细胞可作为研究肝脏疾病、筛选药物代谢和肝毒性的工具。因此，迫切需要进一步研究高效生成肝细胞的方法。从大鼠股骨和胫骨中收集骨髓间充质干细胞（BMSCs）。将FOXA2和HNF1α基因构建到慢病毒载体中，并通过慢病毒介导的过表达系统导入BMSCs。诱导三周后，分析FOXA2和HNF1α以及肝脏特异性基因的表达，并进行肝细胞功能相关检测。FOXA2和HNF1α的过表达诱导BMSCs分化为肝细胞样细胞（HLCs）。通过RT-PCR、Western Blot和免疫荧光检测肝细胞特异性基因和蛋白质。这些HLCs还发挥了一些典型的肝细胞功能，如糖原储存、吲哚菁绿摄取和脂质积累。FOXA2和HNF1α的组合可有效诱导BMSCs分化为HLCs。这是一种在短时间内制备HLCs的新颖且高效的方法。
