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目的探讨肝损伤血清诱导人骨髓间充质干细胞分化为类肝细胞的可行性。方法分离、纯化hMSCs后用含10%肝损伤血清的培养基诱导培养,倒置显微镜连续观察细胞形态变化,并分别于诱导后7、14、21d行免疫细胞化学、糖原染色检测诱导后细胞的功能。结果①MSCs形态均一,呈长梭形;②实验组MSCs在诱导5～7d后细胞形态开始变为不规则圆形、三角形或多角形;③免疫细胞化学:实验组细胞第7天AFP表达呈阳性,第14天AFP表达消失;Alb第14天开始表达,表达量逐渐增强,可持续至第21天;④PAS染色:实验组在诱导21d时细胞有合成糖原的能力,可见胞浆呈红色。结论肝损伤血清可诱导MSCs向类肝细胞分化,为细胞移植治疗肝衰竭提供新的探索思路。
Objective To investigate the feasibility of serum-induced differentiation of human bone marrow mesenchymal stem cells into hepatocyte-like cells by liver injury. Methods The hMSCs were isolated and purified by culture in medium containing 10% serum of liver injury. The morphological changes of cells were observed by inverted microscope. Immunocytochemistry was performed on days 7, 14 and 21 after induction respectively. Glycogen staining Features. Results ① The morphology of MSCs was uniform and long fusiform. ② The morphology of MSCs began to change into irregular circles, triangles or polygons after 5-7 days of induction. ③ Immunocytochemistry: The positive expression of AFP in experimental group on day 7 , The expression of AFP disappeared on the 14th day; Alb began to express on the 14th day, the expression level gradually increased and lasted for 21 days; ④PAS staining: The ability of cells in the experimental group to synthesize glycogen on the 21st day was red. Conclusion Serum of liver injury can induce the differentiation of MSCs into hepatocyte-like cells and provide new exploration ideas for the treatment of liver failure by cell transplantation.