犬骨髓间充质干细胞体外分离培养及分化鉴定

目的:建立犬骨髓间质干细胞(dog bone mesenehymal stem cells,dBMSCs)的体外分离、培养、纯化及鉴定的方法。方法:采用密度梯度离心法结合贴壁筛选法分离d BMSCs,并通过不断传代进行纯化和扩增培养。诱导dBMSCs向成骨、成脂细胞分化,并分别以矿化(钙)结节茜素红染色、油红O染色鉴定。结果:dBMSCs在体外分离培养扩增,形态呈长梭形成纤维细胞样,通过成骨、成脂诱导分化鉴定其为dBMSCs。结论:密度梯度离心法结合贴壁筛选法能有效的获取dBMSCs,所分离培养的d BMSCs同时具有多向分化潜能,是组织工程研究中理想的种子细胞。

Isolation,culture,differentiation and identification of dog mesenehymal stem cells in vivo

Objective:To establish a method for isolation,culture,differentiation and identification of bone mallow mesenchymal stem cells(BMSCs)from dog.Methods:dBMSCs were isolated by combination of gradient centrifugation and adherent wall culture,and induced their differentiation into osteoblasts and adipocytes which were identified by histoehemistry staining and oil red staining.Re-sults:The cultured adherent cells showed spindle and polygonal shape.The identification of dBMSCs was carried out by alizarin red stai-ning after induction of osteoblasts and alkaline phosphatase staining after induction of adipocytes.Conclusion:dBMSCs were isolated and cultured successfully from dog ilium bone marrow by combination of gradient centrifugation and adherent wallculture,which possess multi-directional differentiation potential,is the ideal seed cells in tissue engineering research.