兔骨髓间充质干细胞的体外分离培养、鉴定及向成骨细胞分化诱导  

Isolated Culture,Identification and Induced Differentiation to osteoblast of Rabbit Marrow Mesenchymal Stem Cells in vitro

作　　者：黄旋平[1] 周诺[1] 杨媛媛[1] 江献芳[1] 李华[1] 谢庆条[1] 

机构地区：[1]广西医科大学附属口腔医院口腔颌面外科,广西南宁530021

出　　处：《口腔医学研究》2012年第8期745-748,共4页Journal of Oral Science Research

基　　金：国家自然科学基金资助项目(30760273;30960421);广西壮族自治区自然科学基金资助项目(桂科自0991013Z)

摘　　要：目的:探讨体外分离培养、鉴定兔骨髓间充质干细胞(BMSCs)的方法,并观察其向成骨细胞分化的潜能。方法:采用贴壁分离法分离培养兔BMSCs,通过细胞形态学观察和细胞表面抗原检测对细胞进行鉴定,并向成骨细胞分化诱导培养,通过钙钻法检测碱性磷酸酶表达,茜素红染色观察钙结节形成能力。结果:分离培养3d,可见贴壁细胞呈三角形、成纤维形或多角形外观,培养10～12d细胞长满瓶底,传代后细胞增殖明显加快;使用条件培养基传代培养2周,细胞呈集落生长后开始形成钙结节,茜素红染色阳性。结论:全骨髓贴壁法是一种简单实用的分离培养方法,可获得纯度较高的兔BMSCs,经过诱导后可向成骨细胞分化。Objective： To explore the method of isolated culture and identification of bone marrow mesenchymal stem cells （MSCs） of rabbits in vitro and to investigate the potentiality to osteoblast. Methods; The rabbit＇s MSCs was obtained by adherence separation method and was cultured in the medium for inducing osteoblasts. Then it was identified by cell morphology and cell surface antigen, and its alkaline phosphatase （ALP） expression and potential of calcific nodules was investigated by Gomori method and alizarin red staining, respectively. Result： The wall-adhesive cells demonstrated triangle, long-shuttle or multiangle in shape at the third day for the first washing. After 10 to 12 days＇ culture fibroblastoid cells colonized on the bottom. The BMSCs would grow as colony in conditioned culture medium and then formed calcific nodules positively stained by alizarin red. Conclusion： The adherence separation method is a simple and applicable one for the isolation and culture of whole blood marrow. Highly-purified rabbit MSCs could be isolated using this method and induced into osteoblasts by culture and subculture.

关 键 词：骨髓间充质干细胞 细胞培养 细胞分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]