摘要
目的探索骨髓间充质干细胞（MSCs）体外跨越分化为肝细胞的诱导体系。方法从单核细胞中分离间充质干细胞，用添加20ng／ml HGF、10ng／ml FGF-4、10ng／ml OSM和10％NBS的IMDM培养液诱导间充质干细胞向肝细胞分化。结果经细胞因子诱导的细胞出现肝细胞样变化，随诱导时间延长体积逐渐增大、形态往上皮样转变，胞浆丰富，出现双核或多核细胞。RT—PCR和免疫荧光检测结果显示，AFP在诱导初期表达，在诱导后期表达下降；CK18、ALB和TAT的表达与诱导时间呈线性关系，第20天达到表达高峰；诱导20d后免疫荧光检测，AFP、CK18、ALB和TAT均为阳性；诱导20d细胞的PAS糖原合成反应呈阳性，即具备糖原合成和储存的肝细胞特有功能。结论HGF、FGF4和OSM三种细胞因子组合，可诱导小鼠间充质干细胞向肝细胞分化，该结果为间充质十细胞跨越分化肝细胞的分子机制探讨和临床应用打下了基础。
Objective To establish an effective method for inducing mouse bone marrow mesen chymal stem cells （MSCs） into hepatocytes. Methods Isolated MSCs were selected by plastic adher ence, differentiated in Iscove＇s Modified Dulbecco＇s medium supplemented with 10% new bovine ser um （NBS）, 20 ng/mL hepatocyte growth factor （HGF）, 10 ng/mL fibroblast growth factor 4 （FGF 4） and 10 ng/ml oncostatin m （OSM） for 20 days＇ induction. The medium was changed every 4 days. Results When MSCs were cultured with HGF, FGF 4 and OSM, cuboidal morphology, which was a characteristic of hepatocytes, was observed, and the differentiated ceils also expressed marker genes specific to liver cells in a time-dependent manner, α-fetoprotein （AFP） was expressed on day 10, and CK18, ALB and TAT were detected on day 20, which was in consistent with the immunofluoresence results. Differentiated cells further demonstrated these cells also acquired functional characteristics of hepatocytes of storing glycogen. Conclusion Mouse MSCs can differentiate into hepatocytes when in duced by HGF, FGF-4 and OSM.
出处
《中华肝胆外科杂志》
CAS
CSCD
2008年第10期728-731,共4页
Chinese Journal of Hepatobiliary Surgery
基金
绍兴市科技计划项目（2005141）
