摘要
目的:采用生长因子TGF-β1和bFGF诱导体外培养的兔骨髓间质干细胞(MSCs),转化为韧带样细胞,并研究此种韧带样细胞的生物特性。方法:自幼兔四肢骨抽取骨髓分离纯化MSCs并培养、增殖;采用特定浓度TGF-β1(10ng/ml)和bFGF(25ng/mL)对MSCs进行诱导分化,观察生长因子对MSCs生长、形态的影响,使用MTT法绘制细胞生长曲线,使用天狼腥红染色法定量对比MSCs分泌胶原蛋白量。单纯培养和单一因子诱导组作为对照。结果:TGF-β1和bFGF联合使用组,细胞形态优于空白组及单一因子组,细胞增殖率、胶原分泌量也均高于对照组。结论:联合使用生长因子TGF-β1和bFGF刺激兔MSCs,能够促使兔MSCs定向转化为韧带样细胞,对组织工程前交叉韧带的构建具有积极意义。
Objective： To study the influence of growth factor TGF-β1 and bFGF on inducing the rabbit＇s mesenchymal stem cells （MSC） proliferation and expression in directional differentiation into ligament-like cells in vitro culture. Meanwhile, the biological characteristics of the cells were observed under this condition. Methods： The MSC, which was harvested from baby rabbit＇s bone marrow, was isolated, cultivated, purified and proliferated. Thereafter, a certain amount of growth-factors, TGF-β1 （10 ng/mL） and bFGF（25 ng/mL） were single or combined used to promote directional differentiation in third generation of MSC. The cells growth and morphological features were observed and MTT method was used to measure the cell proliferation with drawing cell growth curve. Picro-Sirius red stain was carried out to quantify the mount of collagen secreting. The simply culture and the one single factor added groups were taken as controls. Results： The cellular morphology, proliferation rate and collagen secretion in TGF-β1 combined bFGF group were significant better or higher than any of control groups. Conclusion： Combined usage of growth factor TGF-β1 with bFGF in a given amount is very helpful for MSC differentiated into ligament-like cell. It is positive findings to select seed cell in ligament tissue engineering.
出处
《现代生物医学进展》
CAS
2013年第24期4638-4642,4662,共6页
Progress in Modern Biomedicine
基金
陕西省社会发展基金课题(2010k13-01-01)
关键词
骨髓间充质细胞
生长因子
组织工程
前交叉韧带
Mesenchymal stem cells（MSC）
Growth factor（GF）
Tissue engineering
Anterior cruciate ligament（ACL）
