兔骨髓间充质干细胞获取与特性的实验研究  

Acquisition and biological characteristics of rabbit bone marrow derived mesenchymal stem cells

作　　者：刘永亮[1] 叶钢[1] 方针强[1] 

机构地区：[1]第三军医大学新桥医院泌尿外科全军肾脏病中心,重庆400037

出　　处：《西北国防医学杂志》2008年第4期247-249,共3页Medical Journal of National Defending Forces in Northwest China

基　　金：重庆市自然基金资助项目(2007BB5022)

摘　　要：目的:完善兔骨髓间充质干细胞(Mesenchymal stem cells,MSCs)体外扩增培养体系并观测其特性。方法:兔骨髓经密度梯度离心获得单核细胞后贴壁培养,光镜、透射电镜观察所获细胞形态,流式细胞术检测细胞周期,体外诱导MSCs向成骨细胞、成脂肪细胞分化并鉴定。结果:原代及传代MSCs为漩涡状贴壁生长的长梭形成纤维细胞样细胞,传代细胞具有类似的生长规律;透射电镜示所获细胞具有较强的自我更新能力;细胞周期分析显示87%以上细胞处于G0/G1期;MSCs经体外诱导可向成骨细胞、成脂肪细胞分化。结论:利用密度梯度离心联合贴壁培养法可分离、纯化MSCs,所获细胞具备成体干细胞特性,于体外可大量扩增,做为种子细胞和载体细胞满足实验需要。Objective：To cultivate rabbit bone marrow derived mesenchymal stem cells （MSCs）in vitro, and find a set of amplification system, then observe its biological characteristics. Methods： The MSCs from rabbit bone marrow were isolated, purified and amplified by density gradient centrifugation and adherent culture in vitro. The changes of MSCs morphology, growth and proliferation were observed by inverted phase contrast microscope , and MSCs uhrastructure and cell cycle were detected by transmission electron microscope and flow cytometer respectively; MSCs were induced into osteogenetic cells or adipose cells and then were verified . Results： The primary cells and the passage cells were mostly fusiform in shape, which was similar to fibroblast cell; during adherence growth, they were arranged tightly in a swirling pattern; the cell cycle analysis showed that more than 87% cells were in G0/G1 phase; after MSCs were induced, cell alkaline phosphatase stain showed that alkaline phosphatase was positive in osteogenetic cell and formed mineralization nodus; oil red O stain showed that red lipid droplet existed in adipose cell. Conclusion ： MSCs can be isolated and cultivated by the method of density gradient centrifugation and adherent culture in vitro, the potentiality of proliferation and multi - directional differentiation can meet the requirements of the further experiments.

关 键 词：细胞培养 间充质干细胞 骨髓 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]