Abstract: Abstract Objective To establish a method of isolation, culture, identification and labeling rabbit mesenchymal stem cells (MSCs) in vitro. Methods MSCs were isolated and cultivated by density gradient and adherent methods. The expressions of CD29 and CD106 of cells were analyzed by using Immunocytochemistry. The third generation of MSCs were labeled by DiI，the labeling efficiency was detected. Results　Primary cultured MSCs adhered to plastic surface within 48h and reached 90% confluence within 7-8 d . MSCs expressed CD29 and CD106. All of the MSCs after labeling by DiI showed red fluorescence by immunofluoroscope. DiI labeling was sensitive and highly efficient to MSCs. Conclusion The method is simple and easy to isolate and cultivate MSCs and it can serve as a routine method. The culturing cells could be used in urethral construction of tissue engineering and then could be used to treat severe hypospadias、hypospadias cripples and urethral stricture in the future.

Key words: bone marrow, mesenehymal stem cells, cell culture