人骨髓间充质干细胞分离培养方法的改进  

Improvement of methods to isolate and culture human mesenchymal stem cells

作　　者：王承云[1] 石磊[2] 夏春[2] 郑欣鹏[3] 张兵[3] 林飞太[3] 王少杰[2] 

机构地区：[1]福建医科大学研究生院,福建省福州市350108 [2]厦门大学附属中山医院关节外科,福建省厦门市361004 [3]厦门大学医学院,福建省厦门市361005

出　　处：《中国组织工程研究与临床康复》2011年第32期5896-5900,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：2009厦门科技基金资助课题子课题(3502Z20094008);课题名称:股骨头坏死间充质干细胞成骨与凋亡过程中NF-kappaB调控caspase-3的研究~~

摘　　要：背景:当前分离培养骨髓间充质干细胞方法的有多种,但在外科手术中进行获取的方法仍然较少,而外科分离所得细胞对于相应疾病的具有揭示原因和提供研究线索的价值。目的:在骨科手术中以直接贴壁差速贴壁分离培养纯化与鉴定骨髓间充质干细胞。方法:在骨科关节置换等大手术中吸取少量人骨髓血,采用直接贴壁法分离骨髓间充质干细胞,在贴壁后36～48h行洗盘处理,并通过长期体外培养,特定培养基自身的筛选作用对细胞进行筛选,当细胞生长达到足量时进行鉴定。结果与结论:采用直接贴壁法能够在长骨髓腔内混合血中分离培养骨髓间充质干细胞并形成集落,其形态学表现和表面分子经鉴定符合骨髓间充质干细胞特点,能够获得符合要求的人骨髓间充质干细胞,从而进行后续实验。BACKGROUND： The methods to isolate mesenchymal stem cells （MSCs） can be classified to be several kinds. However, the method of acquisition in surgery is still less. The isolated MSCs from the patients who receive the replacement operation can offer some data which may reveal the causes or clues of some diseases. OBJECTIVE： To isolate and culture MSCs with an improved technique of differential anchoring velocity and adherence, in order to identify MSCs. METHODS： The bone marrow blood was collected in the femur marrow cavity duiring the hip joint replacement. MSCs were isolated by adherence method in the special medium and the mix medium was flushed 36-48 hours after the inoculation to isolate MSCs from the marrow mixture. In the long-time culture in vitro, the special medium was used to weed other cells out. The anchored cell was cultured and identified when the amount was large enough. RESULTS AND CONCLUSION： MSCs can be isolated from the bone marrow in long cavitas medullaris, and can be cultured into colonies by using adherence method. The morphological features and surface molecules are identified consistent with MSCs, and can obtain standard MSCs, so as to do the follow-up experiment.

关 键 词：人骨髓间充质干细胞 细胞培养 鉴定 流式细胞术 干细胞分离与培养 

分 类 号：R394.2[医药卫生—医学遗传学]