胎肝滤液诱导骨髓间充质干细胞向肝细胞分化

目的 探讨胚胎肝组织滤液定向诱导大鼠骨髓间充质干细胞(BMSCs)分化为肝细胞的诱导条件，为肝组织工程提供新的种子细胞来源。 方法 在体外培养体系中加入胎肝滤液，模拟体内肝脏微环境，诱导BMSCs向肝细胞定向分化，以免疫细胞化学检测肝细胞标志物；PAS检测糖原表达；靛青绿染色检测转化细胞的分化程度；测定细胞培养上清液中丙氨酸氨基转移酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(ALP)的含量以检测其功能状态。 结果 BMSCs经胎肝滤液诱导14d时细胞呈现多角形、卵圆形或圆形细胞的特征性改变；甲胎蛋白(AFP)和白蛋白(ALB)免疫反应、PAS反应和吲哚靛青绿(ICG)摄入实验及上清液中ALT、AST、ALP等酶均在诱导的第7天开始出现，AFP和ALB免疫反应在21d时达高峰；PAS反应和ICG摄入实验随着时间的延长而增强；上清液中的各个酶在14d达高峰，之后呈下降趋势。 结论 胎肝滤液可诱导BMSCs形成具有肝细胞形态和功能特点的细胞。

Fetal rat liver filtrate induces the differentiation of rat bone marrow-derived mesenchymal stem cells into hepatocytes

Objective To explore the possibility that rat bone mesenchymal cells (BMSCs) can differentiate into hepatocytes under the affection of fetal liver filtrate. Methods PAS and green indigo dye were used to detect glycogen and differential level of hepatocytes, respectively. The concentration of ALT, AST, ALP in the culture supernatant were served as markers of hepaocyte function. Results Fourteen days after induced by the fetal liver filtrate, BMSCs changed their shapes into polygon, oval or round. Some of BMSCs were positive for AFP and ALB at 7 days after induction, then the number of positive cells increased, and most of BMSCs expressed AFP and ALB till 21days. The PAS reaction and indocyanine green(ICG) intaking also appeared at 7days. Enzyme in supernatant such as ALT, AST, ALP were fristly detected at 7days and peaked at 14days,then the level declined. Conclusion The fetal rat liver filtrate was able to induce BMSCs into cells with function and characteristics of hepatocytes.