兔骨髓间充质干细胞的分离、培养与鉴定  

APPROACHES TO ISOLATE,CULTURE AND IDENTIFY THE BONE MARROW-DERIVED MESENCHYMAL STEM CELLS OF RABBITS

作　　者：叶发刚[1] 夏长所[2] 张卫兵[3] 夏仁云[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,湖北武汉430030 [2]青岛大学医学院附属医院创伤骨科 [3]南京鼓楼医院骨科

出　　处：《青岛大学医学院学报》2006年第4期330-332,共3页Acta Academiae Medicinae Qingdao Universitatis

摘　　要：目的优化获取兔骨髓间充质干细胞并对其进行纯化、鉴定,同时对其生物学性状进行研究。方法取新生新西兰大耳白兔骨髓5.0 mL,采用密度梯度离心法、贴壁筛选法并结合传代对其进行纯化,观察细胞形态、超微结构和表面标志物的表达,从而对其进行鉴定。观察第1、3、5、8、10代细胞的增殖情况,并绘制出生长曲线。结果经密度梯度离心、贴壁筛选并结合传代所得到的间充质干细胞很纯,第3代和第5代细胞形态单一,细胞排列具有典型的漩涡状结构。第1、3、5代细胞增殖能力强,生长旺盛;而第8、10代细胞增殖明显减弱。所分离培养的细胞表达CD44、CD90,不表达CD34,电镜观察为低分化细胞。结论分离培养的细胞为间充质干细胞且成分单一,具有干细胞特性,第3、5代细胞很纯且其增殖能力强,适用于做以后的研究。Objective To explore the approaches to isolate, culture and identify the bone marrow derived mesenchymal stem cells （BM-MSCs） of rabbits. Methods BM MSCs were isolated from the bone marrow （5.0 mL） of the newborn rabbit via density gradient centrifugation. Light and electron microscopy were used to study the morphologic features and immunocytochemistry to examine the expression of cell surface antigens. The growth curve of cell proliferation was obtained based on the observation of the proliferation status of 1st, 3rd. 5th, 8th, and 10th generation. Results BM-MSCs of 1st, 3rd, 5th generation were very purified and all in the whirlpool-shape. They proliferated more rapidly than those of 8th and 10th generation. These cultured cells showed immunoreactivity to CD44 and CD90 but not CD34 and low-differentiation under the electron microscopy. Conclusion Purified BM-MSCs can be obtained via this protocol. Cells of 1st , 3rd,5th generation with high proliferation are fit to the further experiment.

关 键 词：骨髓间充质干细胞 兔 离心法 梯密度 细胞培养技术 

分 类 号：Q813.11[生物学—生物工程]