体外诱导大鼠骨髓干细胞向肝细胞样细胞的分化

以贴壁细胞分离法分离大鼠骨髓干细胞，体外扩增，采用细胞因子诱导骨髓干细胞定向分化为肝细胞。大鼠骨髓干细胞（MSCs）体外扩增后数量剧增，各诱导组细胞胞浆内可见红染的糖原颗粒；诱导培养4周，细胞上清尿素合成、ALT、AST水平随诱导时间的延长而增加，于第28天达到高峰。Westem—blot检测表明，诱导4d的细胞开始表达AFP蛋白，7d后达到高峰。以FGF-4联合HGF因子组诱导后肝细胞特征最显著。以细胞因子FGF-4、HGF、EGF为主的诱导培养体系能促使骨髓干细胞定向分化为肝细胞，其中FGF-4联合HGF细胞因子的应用更能提高骨髓干细胞定向分化为肝细胞的转化效率。

Differentiation of Rat Bone Mesenchymal Stem Cells into Hepatocytes in Vitro

MSCs are isolated by adherence suspensions in vitro and FGF -4, EGF, HGF are selected as differentiation factors. Western - blot analysis is adopted to detect the differentiated cells function. The differentiated cells are also detected by staining for glycogen and by determining synthesis of BUN, ALT, AST. The number of rat bone MSCs increases sharply after vitro reproduction; in the entire induced groups red glycogen granule can be observed; after 4 weeks induction, the level of AST and ALT in the cultural supernatants increases with the passage of time and reaches the climax on the 28th day, and AFP reaches to pinnacle with the differentiation of MSCs on the 7th cultivation day proved by the Western - blot analysis. The cells induced by FGF - 4 plus HGF appear most obvious hepatocytes character. Through the experiment, we get the conclusion that MSCs can be differentiated into hepatocytes in the differentiation media including HGF, FGF - 4, EGF. Applying FGF - 4 plus HGF is the most effective method of directing MSCs into hepatoeytes.