Isolation, culture and phenotype identification of rat marrow 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 MSCs were isolated and purified from the bone marrow of rat by density gradient centrifugation and by adhering to the culture plastic.After successive subculture and amplification,the growth curve was drawn.The morphology of MSCs was observed in primary and passage culture under inverted microscope.The cells surface antigens of CD29,CD38,CD44 and CD45 were detected through flow cytometer.Results Cultured primary MSCs were spin dle-shaped and had a typical fibroblast-like morphology,proliferated in colonies.After a series of passage,the attached cells became morphological homogeneous and proliferated quickly.The growth curves of passage 1,3 and 5 were quite similar.The surface marker identification showed that CD44 and CD29 were positive,but CD45 andCD38 were negative.Conclusion The MSCs can be successfully isolated,cultured and purified from rat bone marrow through density gradient centrifugation,adherent property and subculture.MSCs can greatly proliferate after a short period of culture in vitro.The MSCs can be used as a potential cell source for tissue engineering. Key words: Bone marrow mesenchymal stem cells, rat;  Cell culturetechnigues;  Phenotype