摘要
目的:探讨兔骨髓间充质干细胞(MSCs)体外分离培养、表型鉴定和标记的方法。方法:采用密度梯度离心法及贴壁分离筛选法分离培养MSCs;采用免疫细胞化学方法检测细胞表面标志抗原CD29和CD106的表达,进行表型鉴定;DiI标记第3代MSCs,观察标记效率。结果:体外培养的原代MSCs48h内可见少量贴壁细胞,7～8d达到90%汇合;免疫细胞化学方法检测细胞表面标志抗原CD29,CD106为阳性;DiI进行细胞标记后,荧光显微镜下见所有MSCs均被标记为红色荧光,敏感性好,标记效率高。结论:此培养方法可以作为培养兔MSCs的常规方法,为构建组织工程尿道提供充足的种子细胞。
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.
出处
《天津医药》
CAS
北大核心
2010年第9期787-789,共3页
Tianjin Medical Journal
基金
国家自然科学基金资助项目(项目编号:30600136)
天津市卫生局科技基金资助项目(项目编号:07KY09)
关键词
骨髓细胞
间质干细胞
细胞
培养的
细胞分离
免疫组织化学
荧光抗体技术
流式细胞术
兔
bone marrow cell mesenchymal stem cells cells
cultured cell separation immunohistochemistry fluorescet antibody technique flow cytometry rabbits
