Repsox对人胚胎干细胞分化为肝细胞的影响

Repsox accelerates the differentiation of human embryonic stem cells toward hepatocytes

目的在肝细胞诱导分化后期加入Repsox,探究其对肝细胞分化效率的影响。方法在人胚胎干细胞定向诱导为肝细胞过程中,从分化第13天开始,在实验组中添加Repsox,对照组中加入等量的DMSO,收集分化至第21天的细胞样品和培养液上清进行检测。通过实时荧光定量聚合酶链反应(RT-qPCR)和酶联免疫吸附(ELISA)测定整个分化过程中TGFβ1的表达量和分泌量变化;测定对照组与实验组细胞中TGFβ1分泌量、白蛋白(albumin)分泌量和尿素(urea)合成量。结果在整个肝细胞分化过程中,TGFβ1先明显上调然后逐渐下调;加入Repsox后,TGFβ1后期下调更为显著。实验组表达白蛋白的细胞数量多于对照组,其白蛋白分泌量以及尿素合成量也明显高于对照组(P<0.05)。结论Repsox能够有效促进人多能干细胞转变为肝细胞。

Objective To investigate the effect of Repsox on the differentiation efficiency of hepatocytes by adding repsox at the later stage of differentiation.Methods In the process of directional induction of human embryonic stem cells(hESCs)into hepatocytes,repsox was added to the experimental group from day 13 and DMSO was added to the control group.Cell samples and supernatant of culture medium were collected at the 21st day of differentiation for detection.The expression of TGFβ1 mRNA and protein during the hepatic differentiation was detected by qRT-PCR and ELISA.The secretion of TGFβ1,albumin and urea were measured.Results In the process of hepatocyte differentiation,TGFβ1 was up-regulated first and then down-regulated gradually;after adding repsox,TGFβ1 was decreased more significantly in the later stage.The number of albumin expressing cells was significantly increased in the Repsox-treated group,and the amount of albumin secretion and urea synthesis in the Repsox-treated group was also significantly higher than that in the control group(P<0.05).Couclusion Repsox can promote the differentiation of human pluripotent stem cells into hepatocytes.