Purification of in vitro cultured adherent rat bone marrow-derived mesenchymal stem cells,.doc

 PAGE \* MERGEFORMAT 9 Purification of in vitro cultured adherent rat bone marrow-derived mesenchymal stem cells, [Abstract] Objective To study the in vitro adhesion separation, purification and proliferation of rat bone marrow-derived mesenchymal stem cells (MSCs), and characterized. Remove method 5 d of Health of the Wistar rat femur and tibia, red to take bone marrow, isolated by screening using separation of adherent rat MSCs, were purified by passage, proliferation, growth curves were measured to detect the growth cycle, flow cytometry surface antigen. Result adherent culture method can be effectively isolated rat MSCs, MSCs uniform spindle morphology, growth curves are S-type, cell cycle showed that about 80.89% the third generation of cells in G0/G1 phase after the third generation cell surface antigen CD90 positive, CD34, CD45 negative. Conclusion of a better in vitro, purification and amplification of rat MSCs methods applied to MSCs for further study. Bone marrow-derived mesenchymal stem cells (MSCs) derived from the mesoderm, with multi-differentiation potential, in different inducing conditions, can become bone cells, cartilage cells, fat cell differentiation, but also has wide variety of sources, easy to culture and transplantation, in vitro gene and high transfection rate, showing in cell and gene therapy a good prospect, has aroused widespread concern in 〔 1〕 . This study was purified rat MSCs in vitro, culture and identification, in order to further explore the application of MSCs provide a basis. 1 Materials and methods 1.1 Materials Clean-class was born 5 d of the Wistar rat (8.0 ~ 10.0 g), purchased from Experimental Animal Center of Jilin University. Main reagents: DMEM medium (GibcoBRL U.S. company), fetal bovine serum (U.S. Hyclone Corporation), rabbit anti-mouse CD90, CD34, CD45 standard direct fluorescent antibody (U.S. BD Inc.), RNA enzyme, propidium iodide (Sigma Company) , 0.25% trypsin, thiazole blue (MTT) and dimethyl sulfoxide