离心分离法和贴壁分离法对培养骨髓间充质干细胞生物学特性影响.docVIP

离心分离法和贴壁分离法对培养骨髓间充质干细胞生物学特性影响 　　[摘要] 目的 探讨分离高质量和高活性骨髓间充质干细胞的方法。方法 选取出生2月龄的健康兔,采用密度梯度离心分离法与贴壁分离法培养骨髓间充质干细胞,检测培养骨髓间充质干细胞生物学特性。结果 贴壁分离法培养的原代骨髓间充质干细胞贴壁增殖快,细胞活性高,而密度梯度离心分离法培养的原代骨髓间充质干细胞贴壁增殖慢,细胞活性低,但这些差别在传代后消失。离心分离法得到的细胞纯度较贴壁分离法获得的细胞纯度高。结论 与离心分离法相比贴壁法获得的原代骨髓间充质干细胞的细胞活性较高、贴壁增殖快,具有方法简单的优点。 　　[关键词] 骨髓; 间充质干细胞; 兔; 细胞培养 　　[中图分类号] R329.2+8 [文献标识码] A [文章编号] 1673-9701(2010)05-29-03 　　 　　Effects of Adherence and Density Gradient Centrifugation Methods on Biological Characteristlcs of Rabbit Marrow Mesenchymal Stem Cells Cultured in Vitro 　　LI Ting1 SUN Jinhu1 ZHAO Liang2 LI Shuai1 　　1.College of Stomatology,Guangxi Medical University,Nanning 530021,China;2.The people’s Hospital of Hechi City,Hechi 546300,China 　　 　　[Abstract] ObjectiveTo study the effects of adherence and density gradient centrifugation(DGC) methods on biological characteristics of rabbit marrow mesenchymal stem cells cultured in vitro. MethodsRabbits aged 2 months were used to isolate marrow mesenchymal stem cells(MSCs). MSCs with adherence and DGC methods were cultured,passaged,amplified and purified in vitro. The living characteristics and adhesive rate of the primary and generations were observed respectively. The growth curve was drawn. The cell cycles and cystoskeleton were tested to study the different MSCs separated methods on the proliferation and metabolism of MSCs. ResultsMSCs were intact and fibroblast cell-shaped. Adhering spindle-shaped cells with higher cytoactive were observed in the adherence and DGC separation groups. The MSCs of primary culture in the adherence group showed more rapid proliferation,earlier colony confluence and shorter time for passage compared with DGC separation group. ConclusionThe MSCs separated by adherence showed a rapid proliferation,earlier colony confluence and shorter time for the first passage. 　　[Key words]Marrow; Mesenchymal stem cells; Rabbit; Cell culture 　　 　　骨髓基质中存在骨髓间充质干细胞(mesenchymal stem cells,MSCs),其具有向成骨细胞、软骨细胞、脂肪细胞、神经细胞等多向分化潜能,是组织工程学技术的重要的种子细胞