人类脐血间充质干细胞在急性肝坏死大鼠体内分化潜能的实验研究

Study on the self-differentation potential of human umbilical cord blood mesenchymal stem cells to liver-like cellsin vivo in rats models suffered from acute hepatic necrosis(AHN)

目的探索人类脐血间充质干细胞在急性肝坏死大鼠体内肝样细胞自主分化的潜能。方法体外分离培养人类脐血间充质干细胞。CC l4橄榄油溶液腹腔注射SD大鼠建立急性肝坏死大鼠模型。造模24 h后肝脏局部注射移植人类脐血间充质干细胞给急性肝坏死大鼠模型,造模后1周、2周和4周分别处死6只模型鼠,取肝脏切片做免疫组织化学检测人类特异性ALB、AFP、HepPar的表达,实时荧光RT-PCR检测人类特异性ALB、AFP、CK19、CK18的表达,测序扩增产物。结果体外分离培养的脐血来源细胞符合人类脐血间充质干细胞的生物学特性。处死移植模型鼠,肝脏免疫组织化学检测显示:人类特异性ALB、AFP、HepPar在接受移植的模型鼠体内呈阳性表达;实时荧光RT-PCR检测显示:人类特异性ALB、AFP、CK19、CK18在接受移植的模型鼠体内呈阳性表达,扩增产物序列与GENEBANK对应序列一致。结论无外部诱导干预,人类脐血间充质干细胞在急性肝坏死大鼠体内已具有肝样细胞自主分化潜能。

Objective To explore the self-differentiation potential of human umbilical cord blood mesenchymal stem cells to liver-like cells in vivo in rats models suffered from acute hepatic necrosis（AHN）.Methods hUCBMSCs were injected into the liver of rats in which AHN was induced by carbon tetrachloride（CCl4）24 h post-molding.AHN rats model was scarified from 1 to 4 weeks post-molding.The transdifferentiation status of the hUCBMSCs was evaluated using immunohistochemistry and real-time RT-PCR.Results The cell line cultured in vitro was in accord with the biological characteristics of human umbilical cord blood mesenchymal stem cell.Immunohistochemistry and real-time PCR analysis showed that transplanted hUCBMSCs expressed several human liver tissue-specific markers（ALB、AFP、CK19、CK18、HepPar）in ratswith AHN.Amplified sequences were corresponding to the same sequence in GENEBANK.Conclusion Human umbilical cord blood mesenchymal stem cells have self-differentiation potential to liver-like cells in vivo in rats models suffered from AHN.