摘要
目的 探讨肝细胞生长因子(Hepatocyte growth factor,HGF)和碱性成纤维细胞生长因子(Basic fibroblast growthfactor,bFGF)诱导大鼠骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BM-MSCs)分化为肝样细胞的可行性。方法取SD大鼠股骨骨髓,直接贴壁法分离纯化BM-MSCs,并体外传代,流式细胞术和成骨诱导对其进行鉴定。取第3代BM-MSCs,分为2组:实验组用HGF(20 ng/ml)和bFGF(10 ng/ml)进行诱导,阴性对照组不加诱导剂,倒置显微镜下观察细胞形态变化;RT-PCR法检测诱导后细胞甲胎蛋白(Alpha fetoprotein,AFP)和白蛋白(Albumin,ALB)基因mRNA的转录水平;免疫细胞化学染色法检测诱导后细胞的AFP和ALB蛋白的表达。结果第3代BM-MSCs表型标志和功能特性均符合MSCs的特点。BM-MSCs经HGF和bFGF诱导后呈肝样细胞形态。实验组细胞可检测出AFP和ALB基因mRNA的表达。实验组细胞诱导后第7天,AFP蛋白开始表达,第14天时表达降低,第21天时不表达;ALB于诱导后第14天出现表达,并随诱导时间的延长表达逐渐增加。结论 HGF和bFGF具有体外诱导BM-MSCs向肝样细胞分化的作用。
Objective To investigate the feasibility of directed differentiation of rat bone marrow mesenchymal stem cells（BM-MSCs）into hepatocyte-like cells with hepatocyte growth facto（rHGF）and basic fibroblast growth factor（bFGF）.Methods BM-MSCs were isolated and purified from rat femora by direct adherence method and subcultured in vitro,then identified by flow cytome-try and ossification induction.The BM-MSCs of passage 3 were induced with HGF（20 ng / ml）＋ bFGF（10 ng / ml）,using those without induction as control,then observed for morphology under invert microscope,determined for transcription levels of alpha feto-protein（AFP）and albumin（ALB）mRNAs by RT-PCR,and for expressions of AFP and ALB by immunocytochemical staining.Re-sults The phenotypic markers and functional characters of BM-MSCs of passage 3 were of characteristics of MSCs.After induction with HGF and bFGF,BM-MSCs showed the morphology of hepatocyte-like cells,in which the expressions of AFP and ALB mRNAs were detected.The expression of AFP was observed on day 7,while decreased on day 14 and disappeared on day 21 after induction.However,the expression of ALB appeared on day 14 and increased with the increasing time for induction.Conclusion HGF and bFGF induced the differentiation of BM-MSCs into hepatocyte-like cells in vitro.
出处
《中国生物制品学杂志》
CAS
CSCD
2011年第1期20-24,共5页
Chinese Journal of Biologicals
关键词
骨髓间充质干细胞
分化
肝细胞生长因子
碱性成纤维细胞生长因子
肝样细胞
Bone marrow mesenchymal stem cells（BM-MSCs）
Differentiation
Hepatocyte growth factor（HGF）
Basic fi-broblast growth factor（bFGF）
Hepatocyte-like cells
作者简介
通讯作者：吴小翎，E-mail：xffcpf@163．com
