Rat bone marrow mesenchymal stem cells isolated and purified by whole bone marrow adherent method and differential passage:A comparison with density gradient centrifugation / 中国组织工程研究

ABSTRACT

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 marrowadherence and isolation of BMSCs in combination of differential passage digestion method.METHODS:By whole bone marrow adherent culturing to isolate and differential digestion and passage of rat BMSCs,the speed ofMSCs in the process of digestion and passage was quicker than other bone marrow cells,as well as the characteristics of adherentspeed,instead of density gradient centrifugation to separate and purify MSCs,and their morphological characteristics wereobserved.Cell growth and proliferation of two kinds of culture method were compared with the density gradient centdfugationseparation.Alkaline phosphatase and oil red staining results were observed to verify BMSC differentiation capacity,to detect thecall surface markers,to validate immune properties and to test its purity.RESULTS AND CONCLUSION:The whole bone marrow adherent culture method isolated and differentially subcultured ratBMSCs.Flow cytometry and osteogenic and adipogenic culture results displayed cytoirnmunity property,purity and differentialcapacity,which have no significant difference compared with density gradient centrifugation.However,cell viability andreproductive activity are obviously elevated.