脐带血单个核细胞体外培养分化为肝细胞的实验观察

Experimental study on the in vitro differentiation of human umbilical cord blood mononuclear cell into hepatocyte

目的探讨脐带血单个核细胞在体外定向分化为肝细胞的条件及能力,为建立脐血干细胞移植治疗慢性肝衰竭提供实验依据。方法采集脐带血分离单个核细胞后,在实验组加入成纤维细胞生长因子(FGF)及促肝细胞生长因子(HGF)、或FGF加胎肝上清液培养,对照组只加FGF培养,观察细胞生长分化状况,免疫组化检测两组人甲胎蛋白(AFP)和白蛋白(Alb)。结果实验组可见类圆形及多边形细胞,呈类肝细胞形态,对照组大多为梭形细胞,未见多边形细胞;实验组细胞AFP、Alb免疫组化染色为阳性,对照组未见AFP,Alb阳性细胞。HGF的诱导分化效果优于胎肝上清液。结论脐带血单个细胞在HGF或胎肝上清液的诱导下可以定向分化为能分泌Alb及AFP的类肝细胞。

Objective To study the condition and potential of human umbilical cord blood monouclear cells （HUCBMNC） to differentiate into hepatocytes in vitro. Methods HUCBMNC were separated from human umbilical cord blood, Fibroblast growth factor （FOF） and hepatocyte growth factor （HGF） or FGF and the supernatant of fetal liver cell culture were added into the HUCBMNC culture flask in the experimental groups. But only FGF was added into the HUCBMNC culture flask in the control group. The expansion and differentiation of HUCBMNC in each group were observed. Human alpha-fetoprotein （AFP） and albumin were detected by immunohistochemistry. Results Most adherent cells were fusiform shape, and no polygon shape cells were found in the control group. These adherent cells were AFP or albumin （ALB） staining negative by immunohistochemistry, thus these adherent cells were most probably fibroblast. However, round shape, spherical and polygon shape adherent cells as well as fusiform cells were found in the experimental group. These adherent cells were AFP or ALB staining positive by immunohistochemistry and therefore they may be hepatocyte. The induction effect of HGF on HUCBMNCs to differentiate into hcpatocytes was better than that of fetal liver cell culture supcrnatant. Conclusion In certain condition, HUCBMNC can differentiate into liver cell in vitro.