体外诱导人骨髓间质干细胞分化为肝细胞样细胞  

Differentiation of human mesenchymal stem cells derived from bone marrow into hepatocyte-like cells in vitro

作　　者：司煜安[1] 许文荣[1] 严永敏[1] 钱晖[1] 朱伟[1] 周忠海[1] 徐静[1] 周洪兴[1] 曹慧玲[1] 李继刚[1] 

机构地区：[1]江苏大学医学技术学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2008年第1期6-10,14,I0001,共7页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(30471938);卫生部科研课题(WKJ2005-2-024);江苏省医学领军人才资助项目;江苏省自然科学基金重点资助项目(BK2007705;BK2007092);镇江市重点实验室资助项目(SH2006066;SH2006070);江苏省高校自然科学基金资助项目(06KJD310040)

摘　　要：目的:探讨人骨髓间质干细胞(MSCs)在体外特定条件下是否可以转化为肝细胞样细胞。方法:分离培养人骨髓间质干细胞,采用肝细胞生长因子(hepatocyte growth factor,HGF)、碱性成纤维细胞生长因子(basic fibrob lastgrowth factor,bFGF)及CC l4损伤的小鼠肝脏共培养等诱导方式,在不同时间点分别用RT-PCR和荧光定量PCR检测肝细胞特异性基因的表达,免疫组织化学染色检测肝细胞标志。结果:在HGF和bFGF诱导第7天时甲胎蛋白(alphafetal prote in,AFP)、细胞角蛋白18(cytokeratin-18,CK18)和色氨酸2,3-双加氧酶(tryptophan 2,3-d ioxygenase,TDO)基因表达阳性,CK18和TDO的表达随诱导时间延长增高;第7天诱导细胞组织化学染色AFP、白蛋白(ALB)、肝细胞核因子(hepatocyte nuc lear factor,HNF)、肝细胞特异性抗原(HSA)和CK18表达阳性。与损伤小鼠肝组织共培养21 d后细胞表达AFP和TDO。结论:HGF和bFGF联合诱导以及与CC l4损伤肝组织共培养的方法可体外促进人MSCs分化为肝细胞样细胞,可为肝脏疾病或肝损伤的干细胞治疗提供细胞来源。Objective： To explore whether human bone marrow mesenchymal stem cells（ MSCs） are able to differentiate into hepatocyte-like ceils under appropriate condition in vitro. The human bone marrow MSCs were isolated and cultured. The MSCs were induced by HGF, bFGF and co-cuhured with damaged mouse liver tissues. The specific genes of hepatocyte were detected by RT-PCR and real-time PCR in different culture period. The hepatic markers were identified by immunofluorescent staining. Results： The expression of AFP, CK-18 and TDO until day 7 were detectable and increasing after prolonged culture. After 7d of inducing, CK18, albumin, AFP, HNF and HSA positive staining was observed in differentiated ceils using immunohistochemistry. AFP and TDO were detected in ceils co-cultured with damaged liver tissues of mouse separated by a transwell membrane for 21 days. Conclusion： Human bone marrow MSCs were able to differentiate into hepatocyte-like ceils and may serve as a source of ceils for cell therapy of he patic diseases.

关 键 词：骨髓间充质干细胞 肝细胞 细胞分化 

分 类 号：R394.2[医药卫生—医学遗传学]