The Culture, Proliferation and Identification of Rat Bone Marrow-Derived Mesenchymal Stem Cells

Objective:To establish a method of isolating and cultivating rat bone marrow mesenchymal stem cells (MSCs) in vitro and the biological characteristics thereof. Methods: Rat MSCs were isolated by density gradient centrifugation and cultured with L-DMEM. The morphology of MSCs was observed in primary and passage culture under inverted microscope. The cell surface antigens of CD29, CD34, CD44 and CD29 were detected through flow cytometer. Results: Cultured primary MSCs were spindle-shaped and had a typical fibroblast-like morphology, proliferated in colonies. After a series of passages, the attached cells became morphological homogeneous and proliferated quickly. During the log phase of growth, cells proliferated with a population doubling time of 45.5h. The surface marker identification showed that CD44 and CD29 were positive, but CD34 and CD45 were negative. Conclusion: The bone marrow MSCs can be successfully isolated, cultured and purified from rat bone marrow through density gradient centrifugation, adherent property and subculture. The MSCs can be used as a potential cell source for tissue engineering.