EPCs在MSCs向肝样细胞转化中作用的初步探讨

The role of endothelial precursor cells in the transformation of mesenchymal stem cells into hepatoid cells

目的:探讨内皮前体细胞(endothelial precursor cells,EPCs)在间充质干细胞(mesenchymal stem cells,MSCs)向肝样细胞转化中的作用。方法:分离培养SD大鼠MSCs和EPCs,取第3代MSCs向肝样细胞转化培养,培养过EPCs的无血清及因子的培养基(endothelial progenitor cell-conditioned medium,EPCs-CM)、无血清的α-最低必需培养基(alpha minimal essential medium,α-MEM)分别与肝细胞转化培养基按1∶1比例配置,作为实验组和模型对照组,正常α-MEM培养基为空白对照组,分别观察细胞形态及数量变化;在培养3、5、7、9d时应用免疫荧光检测甲胎蛋白(alpha fetoprotein,AFP)、清蛋白(albumin,ALB)的表达。结果:空白对照组MSCs形态无明显变化;实验组与模型对照组细胞均出现肝样细胞形态。空白对照组未见AFP、ALB表达,模型对照组和实验组在3、5、7、9d AFP、ALB表达水平均增高,其中实验组比模型对照组AFP、ALB水平增高明显。结论:EPCs在MSCs向肝样细胞转化中可能起一定的促进作用。

Objective:To investigate the role of endothelial precursor cells(EPCs)in the transformation of mesenchymal stem cells(MSCs)into hepatoid cells.Methods:The MSCs and EPCs of SD rats were isolated and cultured,and the third generation of MSCs were taken and cultured into hepatoid cells.The serum-free and factor-free medium(EPCs-CM)and serum-free alpha minimal essential medium(α-MEM)were cultured in 1∶1 ratio to the liver cell transformation medium,as the experimental group and the model control group,respectively.Normalα-MEM medium was used as a blank control group.The changes of cell morphology and number were observed in the above three groups.Immunofluorescence was used to detect the expression of alpha fetoprotein(AFP)and albumin(ALB)at 3,5,7 and 9 d.Results:there was no significant change in MSCs morphology in the blank control group.Both the experimental group and the model control group had hepatoid cell morphology.There was no expression of AFP and ALB in the blank control group,and the expression levels of AFP and ALB in the model control group and the experimental group increased on 3,5,7 and 9 d,and the levels of AFP and ALB in the experimental group were significantly higher than those in the model control group.Conclusion:EPCs may promote the transformation of MSCs into hepatoid cells.