IS OIalion, Culture, ldentification and Labeling of Rabbit Mesenchymal Stem Cells in Vitro

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 was labeled by DiI. The labeling efficiency was detected. Results: The primary cultured MSCs adhered to plastic surface within 48 h and reached 90% confluence within 7-8 d. MSCs expressed CD29 and CD106. All of the MSCs showed red fluorescence by immunofluoroscope after labeling by DiI. 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 could be used to treat severe hypospadias, hypospadias cripples and urethral stricture in the future.