摘要
目的研究碱性成纤维细胞生长因子(bFGF)在体外诱导人骨髓间充质干细胞(MSCs)向心肌样细胞分化中的作用,及其对MSCs增殖的影响。方法用含5-氮杂胞苷(5-aza)的培养基(A组)、含bFGF的培养基(B组)、含5-aza+bFGF的培养基(C组)以及普通培养基(对照组)培养人骨髓MSCs。观察细胞形态的改变;免疫细胞化学法检测α-actin、cTnT和connexin-43的表达;RT-PCR法检测Nkx2·5、GATA-4和cTnT的mRNA水平;MTT法检测MSCs的增殖。结果A组和C组的部分细胞呈肌细胞样改变,表达蛋白α-actin,cTnT和connexin43;A组和C组的cTnT、Nkx2·5和GATA-4的mRNA水平明显高于对照组,B组的Nkx2·5和GATA-4的mRNA水平明显高于对照组;B组细胞增殖明显高于对照组,A组和C组细胞增殖明显低于对照组,但C组明显高于A组。结论bFGF能明显促进MSCs增殖,与5-aza合用能更好地诱导人MSCs向心肌样细胞分化。
Objective To study the potential of bFGF to promote human bone marrow derived MSCs to differentiate into cardiomyogenic cells, and its effect on proliferation of MSCs. Methods MSCs isolated from adult human bone marrow were cultured in four different systems. Group A： medium with 5-aza. Group B： medium with bFGF. Group C ： medium with 5-aza ＋ bFGF. Control group ： medium alone. The morphological changes of MSCs were observed. Then immunocytochemistry staining against α-actin, cTnT, and Connixin43 was performed. The expression of Nkx2. 5, GATA-4 and cTnT was detected by semi-quantitative RT-PCR. The proliferation of MSCs in of Nkx2. 5, GATA.-4 and cTnT was higher than that of control group. In group B, mRNA level of Nkx2. 5 and GATA-4 was higher than that of control group. As compared with the control group, cell proliferation was faster in group B than in group A and C. And the proliferation was faster in group C than in group A. Conclusion bFGF can promote the proliferation of MSCs significantly. When combining with 5-aza, bFGF can promote MSCs to differentiate into cardiomyogenic cells more effectively.
出处
《基础医学与临床》
CSCD
北大核心
2007年第5期514-520,共7页
Basic and Clinical Medicine
基金
北京协和医院2003年重大科研项目
关键词
间充质干细胞
碱性成纤维细胞生长因子
分化
心肌样细胞
mesenchymal stem cells
basic fibroblast growth factor
differentiation
cardiomyogenic cell
