体外诱导大鼠骨髓间充质干细胞定向分化肝样细胞  

Differentiating of Induced Bone Marrow Musenchymal Stem Cells of Rats in Vitro Into Hepatocytes-Like

作　　者：安伟德[1] 胡祥[1] 曹亮[1] 王金晶[2] 李力[2] 宋旭华[2] 刘巨超[2] 徐迎新[2] 

机构地区：[1]大连医科大学第一附属医院普通外科,116011 [2]解放军总医院普通外科研究所

出　　处：《医学研究通讯》2005年第5期10-12,共3页Bulletin of Medical Research

摘　　要：目的观察骨髓间充质干细胞体外诱导分化为肝样细胞。方法自大鼠的股骨中提取骨髓细胞,利用 Percoll 法行骨髓间充质干细胞分离、纯化。取第3代的骨髓间充质干细胞,应用肝细胞生长因子(HGF)20ng/ml 表皮生长因子(EGF)1.5μg/ml 联合诱导。两周后免疫荧光染色及逆转录-多聚酶链反应(RT-PCR)鉴定诱导后细胞白蛋白的 mRNA 的表达。结果诱导分化一周骨髓间充质干细胞变为圆形,两周后免疫荧光染色可见肝细胞标志物 CK18和白蛋白表达,RT-PCR 检测有白蛋白的 mRNA 的表达。结论骨髓间充质干细胞体外诱导能定向分化为肝样细胞。Objective To observe differentiation of inducing of MSCs(bone marrow musenchymal stem cells)of rats in vitro into hepato- cytes-like. Methods Extracting the BMSCs from femur in the rat and using percoll, method realizes BMSCs separation, purification. MSCs of the third generation was cultured by IMDM containing hepatocyte growth factor(HGF)20ng/ml and epidermal growth factor(EGF)1.5μg/ml for 2 weeks. The expressions of CK18 and albumin were detected with immunofluorescence, Albumin mRNA were examined by reverse transcrip- tase polymerase chain reaction(RT-PCR). Results After induced by HGF and EGF for 1 week, the spindle shaped MSCs become round shaped. After induction for 2weeks, the differentiated cells expressed CK18 and albumin were checked. The albumin mRNA expression in differ- entiated cells was checked by RT-PCR. Conclusions MSCs of rats can differentiate into Hepatocytes-like in vitro under the special inducing condition.

关 键 词：细胞定向分化 骨髓间充质干细胞 诱导大鼠 逆转录-多聚酶链反应 Percoll法 RT-PCR检测 免疫荧光染色 肝细胞生长因子 体外诱导分化 表皮生长因子 mRNA 细胞标志物 骨髓细胞 细胞分离 联合诱导 蛋白表达 CK18 白蛋白 第3代 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R295.5[医药卫生—基础医学]