全骨髓直接贴壁分离人骨髓间充质干细胞的生物学特性  

Isolation and charcaterization of human bone marrow mesenchymal stem cells using a whole bone marrow adherent culture technique

作　　者：张颢[1,2] 张斌[2] 程梅[1] 陶艳玲[1] 扈江伟[2] 徐曼[2] 陈虎[2] 

机构地区：[1]哈尔滨市第一医院哈尔滨血液病肿瘤研究所,黑龙江省哈尔滨市150010 [2]解放军军事医学科学院附属医院全军造血干细胞移植中心,北京市100071

出　　处：《中国组织工程研究与临床康复》2012年第1期27-30,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：哈尔滨市科技创新人才研究专项基金项目(2009RFQQS027);课题名称:胎盘间充质干细胞规模化扩增及移植中应用;黑龙江省卫生厅科研课题(2011-518);课题名称:脐带间充质干细胞治疗移植物抗宿主病的临床研究;国家863重大专项课题(2011AA020114);课题名称:干细胞应用于恶性血液病放化疗后造血损伤再生修复的研究及临床评价;首都临床特色应用研究(SQ2010AA0201008009);课题名称:脐带间充质干细胞治疗移植物抗宿主病~~

摘　　要：背景：体外分离培养骨髓间充质干细胞成为实验研究和临床应用的关键环节。目的：采用全骨髓培养法分离人骨髓间充质干细胞,建立一种简单、有效的分离培养骨髓间充质干细胞的方法。方法：通过全骨髓培养法分离培养人骨髓间充质干细胞,并通过传代进行纯化和扩增培养。分别在特定诱导体系中诱导人骨髓间充质干细胞向脂肪、成骨及软骨细胞分化。结果与结论：采用全骨髓培养分离法能获得90%以上纯度的骨髓间充质干细胞;流式细胞仪检测结果显示,贴壁细胞均表达CD73、CD90、CD105,不表达造血细胞表型CD34、CD45和HLA-DR;细胞倍增时间为（24.04±0.49）h;细胞周期分析表明：G0~G1期和S＋G2＋M期所占比例分别为71.63%和28.37%;诱导分化结果显示,人骨髓间充质干细胞能够向脂肪、骨和软骨细胞分化。说明全骨髓分离培养法是一种较好的分离人骨髓间充质干细胞的方法。BACKGROUND： Bone marrow mesenchymal stem cells （BMSCs） are widely used in cell therapy and tissue regeneration, so their isolation and culture are a key technique for research and clinical application. OBJECTIVE： To isolate the BMSCs by whole bone marrow adherent culture technique and to establish an easy and effective method to isolate and culture BMSCs. METHODS： The BMSCs were isolated by whole bone marrow adherent culture technique, and purified and amplified by passage. BMSCs were induced to differentiate into adipocytes, osteocytes and chondrocytes in special differentiation condition. RESULTS AND CONCLUTION： The purity of the BMSCs isolated by whole bone marrow adherent culture technique was more than 90%. The adherent cells displayed an abundant presence of CD73, CD90, CD105 and absence of Hematopoietic cell phenotype CD34, CD45 and HLA-DR which were detected by flow cytometry. The cell doubling time was （24.04±0.49） hours; Cell cycle showed that there was percentage of stem cells as G0 /G1 71.63% and S＋G2＋M 28.37% respectively. The induced and differentiated results showed than BMSCs could be differentiated into adipocytes, osteocytes and chondrocytes. Based on these findings, the whole bone marrow adherent culture technique is a better method to obtain MSCs from human bone marrow.

关 键 词：全骨髓培养 间充质干细胞 分离 培养 分化 

分 类 号：R394.2[医药卫生—医学遗传学]