Objective To research the methods and techniques of SD rat bone marrow stromal cells （MSCs）culture in vitro and to provide a large number of MSCs for cell therapy. Methods Bone marrow from the femur and tibia of the early age SD rats was taken to culture to the passage 1-4 （P1-P4）, its growth was observed and P3 cells were evaluated by HE staining and immunohistochemisty. Results The growing speed of P1-P3 was faster than that of P0, cells fusion was 85%-90% after 3-4 d and the cells were arranged in groups liked whirlpool shape or parallel; （3.4-3.6）×104/cm2 cells were gained and the total cell number of P1-P3 was 4.08×106, 2.44×107 and 2.85×108 respectively, the rate of trypan blue rejecting stained was 95%-97%. P4’s growing speed was slower than before, 1.42×109 in 3.0×104/cm2 cells were gained, and the rate of rejecting stained was 95%. P4-cell output was amplified nearly 2 000-fold higher than P0-cell. P3 immunohistochemical analysis indicated CD105+ cells 61.9%， CD44+ 45.4%， CD29+ 16.8%， CD45+ 8.2%， CD31+ 13.7%， CD34+ 8.3% and CD11b+ 1.5%, respectively. Conclusion The culture of whole bone marrow is suitable for a large number of MSCs provision in vitro, and can meet the needs of the cell therapy research.