玻璃粘连蛋白对大鼠骨髓间充质干细胞成骨分化影响

EFFECT OF VITRONECTIN ON OSTEOGENIC DIFFERENTIATION OF MESENCHYMAL STEM CELLS IN RATS

目的体外分离纯化培养大鼠骨髓间充质干细胞(rMSC),研究玻璃粘连蛋白(VN)在rMSC向成骨方向分化过程中的作用。方法贴壁分离培养法分离纯化rMSC,观察第1、3、5、8、10代细胞的增殖情况,并绘制出生长曲线。选第3代细胞用以VN包被的培养板培养,每4d检测碱性磷酸酶活性及钙结节的形成。结果经贴壁筛选法分离的rMSC生长曲线呈S形,第3、4、5代细胞排列具有典型的漩涡状结构;细胞表达CD44、CD90,而不表达CD34。用以VN包被的培养板培养12d,碱性磷酸酶染色呈阳性,硝酸银染色显示有钙结节形成。结论VN有诱导rMSC向成骨方向分化的作用。

Objective To isolate, purify and cultivate the rat mesenehymal stem cells （rMSCs） in vitro, and study the effect of vitroneetin （VN） on osteogenie differentiation of rMSCs. Methods rMSCs were isolated and purified by wall adhesion culture procedure in vitro. The growth curve of cell proliferation was obtained based on the observation of the proliferation status of 1st, 3rd, 5th, 8th, and 10th generation. The cells of the 3rd generation were cultured in culture plates coated by VN, and calci fying nodule and alkaline phosphatase （ALP） activity checked every four days. Results The rMSCs growth curve appearred as S form, and 3rd,4th and 5th generation manifested as typical whirlpool structure. These cells expressed CD44 and CD90, but not CD34. The cells cultured for 12 days in eultureplates and coated with VN showed positive ALP expression and calcifying nodules. Conclusion Vitronectin pocesses a potential of inducing RMSC towards osteogenic differentiation.