Isolation,cultivation and identification of mesenchymal stem cells of rats in vitro

Objective To establish a simple and effective method of isolation,purification and culture of rat bone marrow mesenchymal stem cells(MSC) in vitro,and to explore its biological characteristics and identify the cells in order to provide evidence for gastric epithelial cells differentiation.Methods MSCs of Sprague Dawley(SD) rat were isolated,purified,subcultured and amplified by means of adherence culture.Cell growth was observed under an inverted microscope and membrane antigen CD29,CD44,CD34 were detected through immunohistochemistry.Osteoblastic differentiation and adipocytic differentiation were induced by different inducers for 2～3 weeks in vitro.Morphology changes were observed and MSCs were stained with oil red O after 3 weeks to verify the results of differentiation.Results MSCs could be separated 8～10 days after primary culture and morphology of MSCs was identical fibroblast-like.MSCs were passaged every 3～4 days.Immunohistochemistry staining showed that CD29,CD44 were positive and CD34 was negative in more than 90% MSCs.They could be induced to differentiate into adipoctyes and osteoblasts.Conclusion Adherence culture is a simple and practical method for separated and amplified MSCs.