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首页> 外文期刊>Cytotechnology >Evaluation of isolation methods and culture conditions for rat bone marrow mesenchymal stem cells
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Evaluation of isolation methods and culture conditions for rat bone marrow mesenchymal stem cells

机译：大鼠骨髓间充质干细胞分离方法及培养条件的评价

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Bone marrow mesenchymal stem cells (bMSCs) are multipotent and preferred for cell therapy. However, the content of bMSCs is very low. To propagate a large number of primary bMSCs rapidly has become a prerequisite for bMSC study and application. Different methods of isolating and culturing bMSC were used and compared among groups: bMSCs of group A are isolated using direct adherence method and cultured by conventional medium changing; of group B are isolated using direct adherence method and cultured by low volume medium changing; of group C are isolated using density gradient centrifugation and cultured by conventional medium changing; of group D are isolated using density gradient centrifugation and cultured by low volume medium changing. The average population doubling time (PDT), average generation time and the cumulative cell doubling level were calculated for every group. bMSCs cultured with complete medium containing 10, 11 and 15?% FBS were allocated into group a, b and c separatedly. Cell numbers were counted everyday under a microscope, the population doubling level curve was plotted and PDT was calculated. The growth curve of bMSC in group a, b and c was made. Both density gradient centrifugation and direct adherence methods obtained relatively pure bMSCs. A larger quantity of primary bMSCs were obtained by direct adherence. bMSC proliferation was faster when cultured via the low volume medium changing method at a serum concentration of 11?% than the other methods. Isolating bMSC by direct adherence and culturing by low volume medium changing at a serum concentration of 11?% is preferential for bMSC propagation.
机译：骨髓间充质干细胞（bMSCs）具有多能优势，是细胞治疗的首选。但是，bMSC的含量很低。快速繁殖大量原代bMSC已成为bMSC研究和应用的先决条件。 bMSCs的分离培养方法不同，各组之间进行比较：A组bMSCs采用直接贴壁法分离，常规换培养。 B组采用直接贴壁法分离，通过低体积培养基更换培养。使用密度梯度离心法分离C组中的一部分，并通过常规培养基更换进行培养。使用密度梯度离心法分离D组的D，并通过低体积的培养基更换进行培养。计算每组的平均群体倍增时间（PDT），平均生成时间和累积细胞倍增水平。用含有10％，11％和15％FBS的完全培养基培养的bMSC分别分为a，b和c组。每天在显微镜下计数细胞数，绘制群体倍增水平曲线并计算PDT。绘制了a，b和c组bMSC的生长曲线。密度梯度离心法和直接贴附法均获得了相对纯的bMSC。通过直接粘附获得了大量的原代bMSC。通过低容量培养基更换法在血清浓度为11％的条件下培养时，bMSC的增殖比其他方法快。通过直接粘附分离bMSC并通过以11％的血清浓度改变的小体积培养基进行培养是bMSC繁殖的优先选择。

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