全骨髓贴壁并差速传代分离纯化大鼠骨髓间充质干细胞:与密度梯度离心法的比较

背景:目前国内外分离纯化骨髓间充质干细胞有两种主要方法;密度梯度离心法及全骨髓贴壁法,前者步骤较复杂,后者简单易操作,但纯化效果不理想.目的:在全骨髓贴壁分离骨髓间充质干细胞基础上,并用差速传代消化法,建立大鼠骨髓间充质干细胞体外分离培养纯化方法.方法:全骨髓贴壁培养法分离并差速消化传代大鼠骨髓间充质干细胞,利用间充质干细胞在消化传代过程中较其他骨髓细胞消化悬浮速度快,以及贴壁快的特点,代替密度梯度离心操作来分离纯化间充质干细胞,对其形态学特征进行观察,并与密度梯度离心法比较两种分离培养法的细胞生长增殖情况;观察碱性磷酸酶及油红染色情况,验证骨髓间充质干细胞的分化能力;检测细胞表面标记物,验证免疫特性及检测其纯度.结果与结论:全骨髓贴壁培养法分离并差速传代大鼠骨髓间充质干细胞,流式细胞鉴定、成骨成脂肪培养结果显示其细胞免疫特性、纯度、分化能力与密度梯度离心法无显著差异,但细胞活力,增殖能力有明显提高.

Rat bone marrow mesenchymal stem cells isolated and purified by whole bone marrow adherent method and differential passage:A comparison with density gradient centrifugation

BACKGROUND:At present,there are two main methods of isolating and purifying bone marrow mesenchymal stem cells(BMSCs):density gradient centrifugation and the whole bone marrow adherent method.The former has complicated procedures,and the latter has simple operation,but the purified outcomes are not ideal.OBJECTIVE:To establish a rat BMSCs isolated and cultured in vitro purification methods on the basis of the whole bone marrow adherence and isolation of BMSCs in combination of differential passage digestion...