Compare Of Different Culture Methods For Rat Bone Marrow Mesenchymal Stem Cell And Cultured Cell Identification

Objective: To study the rat bone marrow mesenchymal stem cells (Bone, marrowmesenchymal stem cell BMSCs), the differences of the different training methods, and thecells do morphological observation and identification of surface markers on the trainalgebra. To experiment in the future to provide high purity BMSCs provide a method andbasis. Methods:150g~180g about two male SD rats hind femur and tibia bone marrow,respectively, total bone marrow culture method and the density-density gradientcentrifugation culture. Two kinds of isolated cells, the screening of adherent cells andcultured easily attached to the plastic products of the characteristics of BMSCs was givenseveral passages purification. Observation are two ways to cultured cells of differentalgebraic form, draw adherent cell growth curve, P5generation, flow cytometry, CD34,CD29, CD44, CD45cytokines, identification of bone marrow mesenchymal stem cells.Result: Two kinds of methods for cell fusiform, showing characteristic swirling growthvisible on the morphological observation. Draw two kinds of methods P0to P10generation of cell growth curve, the number of passages in each generation of single dishfor the detection of indicators for single-sample, paired T-test analysis: P＜0.05. Provethat the whole bone marrow culture faster cell proliferation. P3on behalf of two groups ofcells for flow cytometry results of CD34, CD45negative, CD29, CD44-positive. Provetwo kinds of methods in cultured cells in line with the characteristics of BMSCs.Conclusion: bone marrow culture compared to the density-density gradient centrifugationmethod can not only successfully isolated bone marrow mesenchymal stem cells and thereagent requirements, more economical, easier to operate, easy, and training required forthe shorter cycle times. Whole bone marrow culture of BMSCs cultured and in the5~10 generation of cell purity, activity and BMSCs purity is more stable, can be BMSCsinduced more differentiation and application of BMSCs animal studies and clinicalapplications.