大鼠骨髓间充质干细胞体外分离培养及成骨成脂诱导

目的建立大鼠骨髓间充质干细胞（BMSCs）体外分离、纯化、扩增的方法及诱导BMSCs成骨成脂分化,提供理想的组织工程种子细胞。方法采用贴壁筛选法分离BMSCs,通过不断传代进行纯化和扩增培养,并绘制细胞生长曲线。诱导后检测油红O染色和矿化结节。结果 BMSCs在体外分离培养扩增,细胞形态为长梭形,流式检测分析P3和P5细胞表面标记物CD44、CD90为阳性,CD45为阴性。细胞生长曲线呈S形。经成脂诱导培养,脂肪的特异性油红O染色为阳性;经成骨诱导培养,形成了矿化结节。结论本实验分离培养的细胞具有BMSCs的表面标记特征和诱导分化为脂肪细胞和成骨细胞的潜能,为BMSCs在组织工程中的应用提供基础理论和实验依据。

Isolation,Cultivation and Differentiation of Rat Bone Mesenchymal Stem Cells into Adipocytes and Osteoblasts in Vitro

Objective To establish the methods of isolation,purification and amplication of rat bone mesenchymal stem cells（BMSCs） in vitro and induce osteogenic and adipogenic differentiation of BMSCs. Methods BMSCs were isolated and purified by adhesion culture procedure and passaged in vitro,and the cell growth curve was drawn.P3-BMSCs were stained by Oil Red O and Alizarin Red S after induction.Results BMSCs had typical feature of spindle-shape and whirlpool arrangement in culture.The result of flow cytometer showed that the BMSCs in generation 3 and 5 expressed CD44,CD90,but did not express CD45.The growth curve appearred as S form,and Oil Red O and Alizarin Red S staining was positive.Conclusion Cells cultured in this method possess the character of surface antigens of MSCs,and can be induced to differentiate into osteogenic cells and adipogenic cells.