犬骨髓间质干细胞体外培养和鉴定  

Isolation and identification of canine bone marrow mesenchymal stem cell

作　　者：曾晶晶[1] 周诺[1] 黄旋平[1] 廖凤春[1] 陈亚莉[1] 

机构地区：[1]广西医科大学附属口腔医院,南宁市530021

出　　处：《实用医学杂志》2013年第16期2649-2651,共3页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:81160132);广西壮族自治区自然科学基金资助项目(编号:桂科自0991013Z)

摘　　要：目的:建立犬骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的体外分离、培养和鉴定方法,以期为应用BMSCs作为种子细胞提供实验基础。方法:采集犬胫骨骨髓组织,采用全骨髓贴壁培养法体外分离、培养和扩增犬骨髓间充质干细胞。倒置相差显微镜观察细胞形态,流式细胞术检测细胞表面标志物,成骨和成软骨诱导培养鉴定,观察细胞生物学特性。结果:原代和传代的BMSCs贴壁生长,呈纺锤形、长梭形,漩涡状排列、成集落生长。细胞表面标志物CD 90、CD 29、CD44分子阳性,CD34和HLA-DR分子阴性。经成骨诱导后钙结节形成并茜素红染色阳性,诱导期间碱性磷酸酶活性持续增高。成软骨诱导后HE染色见软骨组织,甲苯胺蓝染色见细胞基质异染成紫红色。结论:成功建立了犬BMSCs体外分离纯化、培养的有效方法,扩增的BMSCs具有多向分化潜能,是理想的组织工程种子细胞。Objective To establish methods for isolation and identification of canine bone marrow mesenchymal stem cells （BMSCs） in vitro. Methods Bone marrow was aspirated from dog tibias, and the BMSCs＇s were isolated by adherence method. The BMSCs were identified by cell morphology and cell surface antigens. They were also induced to differentiate towards osteoblasts and chondroblasts, which were identified by corresponding histochemistry staining. Result The primary and passaged BMSCs were spindle-shaped with adhesion characters, and grew in colonies. These cells were positive for essential MSC surface molecules including CD90, CD44 and CD29, and were negative for most haematopoietic markers, such asCD34 and HLA-DR. The BMSCs could be differentiated towards osteogenic and chondrogenic phenotypes. Conclusion The adherence method is a simple and feasible method for canine BMSCs isolation. BMSCs posses multidirectional differentiation potentials, which may be an ideal seed cells for tissue engineering.

关 键 词：骨髓 间充质干细胞 犬 细胞培养 诱导分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]