两种分离方法培养成体大鼠骨髓间充质干细胞的比较分析

Comparative Analysis of Adherent Culture Method and Density Gradient Centrifugation in Culturing Adult Rat Bone Marrow Mesenchymal Stem Cells

目的比较分析目前较简单易行的贴壁培养法和密度梯度离心法培养大鼠骨髓间充质干细胞(MSCs)的效果.方法用贴壁培养法和密度梯度离心法分别培养大鼠MSCs,流式细胞仪检测MSCs的表面标记物.结果贴壁培养法培养的原代细胞较密度梯度离心法培养的原代细胞生长快,但传代细胞这一差异不明显;而密度梯度离心法培养的原代细胞较贴壁培养法培养的原代细胞形态一致,随着传代的增加,贴壁培养法培养的细胞形态逐步趋于一致.流式细胞仪检测示:贴壁培养法培养的细胞:CD29(99.8%)、CD90(99.6%)、CD34(3.01%);密度梯度离心法培养的细胞:CD29(100%)、CD90(97.1%)、CD34(0.89%).结论用贴壁培养法和密度梯度离心法都可以得到纯度较高的MSCs,两种培养方法的效果无明显差异.

Objective To compare and analyze the results of culturing mesenchymal stem cells（MSCs） by adherent culture method and density gradient centrifugation.Methods Rat bone marrow MSCs were isolated and cultured by adherent culture method and density gradient centrifugation separately.Flow cytometry was used to detect their surface markers.Results The primary cultured cells by adherent culture method grew faster than those cells by density gradient centrifugation,but the difference was not obvious in the passage cells.The shape of primary cultured cells by density gradient centrifugation was similar to these cells by adherent culture method,but with the passaging,cultured cells in adherent culture gradually form into cell line.The tests of flow cytometry showed CD29（99.8%）,CD90（99.6%）,and CD34（3.01%）in these cells by adherent culture method.And in these cells by density gradient centrifugation,the results showed CD29（100%）,CD90（97.1%）,and CD34（0.89%）.Conclusion Rat bone marrow MSCs can be obtained by adherent culture method and density gradient centrifugation,and there is no significant difference between the two culture ways.