肝纤维化大鼠骨髓间充质干细胞体外诱导为类肝细胞  

Bone marrow mesenchymal stem cells from rats with hepatic fibrosis differentiate into hepatocyte-like cells in vitro

作　　者：何文艳[1] 刘静[2] 刘树贤[2] 

机构地区：[1]河北医科大学石油临床医学院感染科,河北廊坊065000 [2]河北医科大学第一医院肝病中心实验室,河北石家庄050031

出　　处：《基础医学与临床》2009年第3期277-282,共6页Basic and Clinical Medicine

基　　金：河北省卫生厅科研课题(062761355)

摘　　要：目的观察正常、肝纤维化模型组大鼠骨髓间充质干细胞(MSCs)体外诱导分化为类肝细胞的差异。方法Wistar大鼠随机分为对照组和肝纤维化模型组。用皮下注射CCl4乳剂建立肝纤维化模型。用密度梯度离心法和贴壁法分离大鼠的MSCs,经培养传代获得纯化的MSCs。用肝细胞生长因子(HGF)和成纤维细胞生长因子-4(FGF-4)诱导培养纯化的MSCs。留取15、21和27 d细胞培养液进行白蛋白(Alb)、甲胎蛋白(AFP)检测;于27 d收集细胞爬片,进行糖原染色和免疫细胞化学染色检测MSCs中CK-18、CK-19。结果于诱导15、21和27 d,2组经诱导培养的MSCs AFP水平均高于未诱导的MSCs(P<0.01),其中21 d AFP水平最高;而白蛋白水平21和27 d 2组经诱导培养的MSCs均高于未诱导的MSCs(P<0.01),15 d无差异,27 d最高。27 d 2组经诱导培养的MSCs糖原染色和CK-18、CK-19免疫细胞化学染色均阳性,未诱导的MSCs糖原染色、CK-18、CK-19均阴性。从AFP、白蛋白水平综合比较,正常和肝纤维化模型组大鼠MSCs诱导效果无明显差别。结论HGF、FGF-4可在体外诱导肝纤维化大鼠的MSCs分化为具有肝细胞样细胞表型和功能的类肝细胞。Objective To investigate the difference of differentiation from rat bone marrow mesenchymal stem ceils （MSCs） from normal group and hepatic fibrosis model group into hepatocyte-like cells. Methods Wistar rats were randomly divided into two groups ： normal group and hepatic fibrosis model group. The liver fibrosis was induced by CCL4. MSCs were isolated by combining gradient density centrifugation with plastic adherence. Pure MSCs were obtained by cultivation and passage. The cells then treated with HGF and FGF-4. Levels of AFP and albumin from supernatant were determined on day 15, 21 and 27. On day 27, cells of induced and non-induced were collected, glycogen store of hepatocytes and the expressions of CK-18 and CK-19 were detected. Results The level of AFP in induced MSCs was higher on day 15, 21, 27, and reached the peak on day 21 ; there was no significant difference between induced and non-induced MSCs in albumin levels on day 15, but on day 21,27, compared with the noninduced MSCs, the albumin level in the induced MSCs was higher and reaches its peak on day 27 ; glycogen storage of induced MSCs was measured on day 27 as compared with non-induced MSCs; the induced MSCs expressed CK- 18 and CK-19 while the non-induced MSCs did not. Compared by the levels of AFP and albumin, there was no significant difference in differentiation effect of MSCs between the normal group and hepatic fibrosis model group. Conclusion Rat MSCs of hepatic fibrosis model group could differentiate into hepatocyte-like cells with hepatic phenotype and biological function in the presence HGF and FGF-4.

关 键 词：肝纤维化 骨髓间充质干细胞 类肝细胞 大鼠 

分 类 号：R575.2[医药卫生—消化系统]