成人血源性间充质干细胞的分离和鉴定  

Isolation and Identification of Adult Blood-Derived Mesenchymal Stem Cells

作　　者：曹聪[1] 董英海[1] 董宇启[1] 崔磊[2] 刘伟[2] 曹谊林[2] 

机构地区：[1]上海第二医科大学仁济医院骨科,上海200127 [2]上海第二医科大学第九人民医院整复外科

出　　处：《上海第二医科大学学报》2005年第7期661-665,共5页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：上海市科委重点攻关项目(ODJ14001-7);上海第二医科大学自然科学基金(2001校14)资助项目.

摘　　要：目的建立一种分离、培养扩增成人血源性间充质干细胞(MSCs)的方法进行鉴定,并与骨髓源性MSCs比较生物学特性。方法30名成年志愿者随机平分为二组,一组直接抽静脉血,并细分为全血细胞法组和外周血密度梯度离心法组,同时抽骨髓为骨髓密度梯度离心法组;另一组为血细胞分离机法组。各组进行细胞存活率、集落形成率、形态学、流式表型分析、胶原染色等研究。结果外周血密度梯度离心法分离成人静脉血后培养,贴壁细胞呈梭形,集落形成率为(0.12±0.08)/106单个核细胞,传代扩增到第5代时每份平均细胞数达51.4674×106,表达CD44、CD54、CD105和CD166,不表达CD14、CD34、CD45和CD31,细胞分泌Ⅰ、Ⅲ型胶原。全血细胞法和血细胞分离机法获得的细胞不能连续多次传代。结论外周血密度梯度离心法比全血细胞法和血细胞分离机法简单,贴壁细胞培养扩增容易,获得的血源性MSCs与骨髓源性MSCs的生物学特性相似。ObjectiveTo isolate and identify the mesenchymal stem cells(MSCs) from human adult peripheral blood.MethodsBlood samples were collected from adult individuls and they were divided into whole blood group(n=15), density gradient centrifugation group(n=15) and blood separator group(n=15). MSCs were identified according to morphology, cell-surface antigen profile, and production of functional proteins.ResultsThe adult blood-derived MSCs separated by density gradient centrifugation grew best. In vitro cultivation, the expressed phenotype of adherent spindle-shaped cells was CD_(44)(+), CD_(54)(+), CD_(105)(+), CD_(166)(+), but CD_(14)(-), CD_(34)(-), CD_(45)(-), CD_(31)(-). They also producted collagen type and .ConclusionDensity gradient centrifugation is the best method of isolating MSCs from human adult peripheral blood. The biological characteristics of the peripheral blood-derived MSCs resemble those of the bone marrow-derived MSCs.

关 键 词：血细胞 细胞培养 间充质干细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]