肝再生大鼠血清诱导骨髓干细胞向肝细胞分化的实验研究

Effects of hepatectomized rat serum on the transdifferentiation of adult rat bone marrow cells into hepatocyte-like cells

目的 观察肝大部切除大鼠再生血清和肝细胞生长因子(HGF)诱导骨髓干细胞向肝实质细胞分化的作用;探讨骨髓干细胞向肝细胞分化和增殖的体外培养条件和方法,为患者应用自身骨髓细胞修复损伤肝脏提供实验依据。 方法 制作肝大部切除大鼠肝再生动物模型,收集术后24 h血清;分别应用鼠肝再生血清和HGF对大鼠骨髓细胞进行定向诱导分化培养;以免疫组化、RT-PCR和western blot等方法对分化不同阶段细胞进行检测。将分化细胞经尾静脉输入同系大鼠,观察输注细胞在肝、脾内的生长情况。 结果 活体移植分化细胞能定向迁移至肝和脾体外及体内分化细胞在mRNA水平和蛋白质水平均表达白蛋白,并在一定时期内表达甲胎蛋白。 结论 大鼠骨髓干细胞在肝大部切除再生血清或HGF的诱导下能横向分化为肝实质样细胞。

Objective To study the effects of hepatectomized rat serum and hepatocyte growth factor (HGF) on the transdifferentiation of adult rat bone marrow stem cells (ABMSCs) into hepatic parenchymal cells. Methods The serum was collected from the rats 24 hours after being subjected to subtotal hepatectomy. ABMSCs were collected and cultured in DMEM/F12 (1:1) containing the hepaetectomized rat serum or HGF. The differentiated hepatocyte-like cells were labeled with CM-DiI and administrated by tail vein injection into the isogeneic rats. The cultured and injected cells were both identified by immunocytochemistry and cultured cells were assayed using RT-PCR and Western blot. Results Hepatectomized rat serum and HGF were demonstrated to have the effect of inducing transdifferentiation of ABMSCs into hepatocyte-like cells in vitro. The differentiated cells expressed albumin mRNA and albumin after 7 days++'s co-incubation. Albumin-expressing and CM-DiI positive hepatocyte-like cells were characterized in livers and spleens of the rats injected with the cultivated cells. Conclusion ABMSCs could transdifferentiate into hepatic parenchymal cells by hepatectomized rat serum or HGF.