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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 induce rabbit bone marrow mesenchymal stem cells (MSCs) cultured in vitro with growth factors TGF-β1 and bFGF to transform them into ligament-like cells and to study the biological characteristics of such ligament-like cells. Methods: MSCs were isolated and purified from bone marrow of extremities of young rabbits. MSCs were induced to differentiate by specific concentration of TGF-β1 (10ng / ml) and bFGF (25ng / mL), and the effects of growth factors on the growth and morphology of MSCs The MTT method was used to draw the cell growth curve. The amount of collagen secreted by MSCs was quantitatively compared with Sirius bright red staining. Simple culture and single factor induction group served as controls. Results: The combination of TGF-β1 and bFGF had better cell morphology and cell proliferation rate and collagen secretion compared with the blank group and the single factor group. Conclusion: The combined use of growth factors TGF-β1 and bFGF to stimulate rabbit MSCs can promote the MSCs directional transformation into ligament-like cells, which has positive significance for the construction of anterior cruciate ligament.