人脐血源性间充质干细胞的体外培养及生物学鉴定  

Culture and identification of mesenchymal stem cells derived from human umbilical blood

作　　者：陈镭[1] 苗宗宁[2] 张东强[3] 陆华[2] 王玲[2] 栾文忠[4] 惠国桢[5] 

机构地区：[1]天津医科大学总医院神经外科,300070 [2]无锡市第三人民医院细胞室 [3]沈阳市第五人民医院神经外科 [4]北京大学人民医院神经外科 [5]苏州大学第一附属医院神经外科

出　　处：《江苏医药》2005年第7期481-483,共3页Jiangsu Medical Journal

基　　金：国家自然科学基金资助(30271325);江苏省自然基金资助(BK2001170)

摘　　要：目的建立体外培养、扩增人脐带血间充质干细胞(MSCs)的方法,初步鉴定其生物学特性。方法无菌条件下取正常足月剖宫产的脐带血,用淋巴细胞分离液分离脐血的单个核细胞,以偏酸性的MesencultTM作为培养基进行培养和纯化获得贴壁细胞层,测定MSCs的生长曲线,用流式细胞技术分析细胞的表面抗原。结果来源于脐血的单个核细胞在体外用合适的培养基培养,细胞贴壁后出现破骨样及间充质样的细胞。间充质细胞为成纤维样的细胞形态,并表达MSCs相关的抗原CD29、CD44、CD105,但不表达CD34、CD45、CD106和HLADR。这与源于骨髓的MSCs一致。结论脐带血中含有的MSCs可在体外培养、扩增,能够为实验和临床的应用提供一种新的间充质干细胞来源。Objective To establish the method of in vitro culture and expansion of human umbilical blood-derived mesenchymal stem cells (MSCs) and to investigate their biological characteristics. Methods Human umbilical cord blood (HUCB) was collected from full term deliveries scheduled for cesarean section, the cord blood mononuclear cells were isolated by lymphocyte separation medium, purified and cultured with Mesencult TM medium and acidic environment to produce adherent layer. Growth curves of MSCs were drawn and surface antigenic features were analyzed by flow cytometry technique. Results When HUCB-derived mononuclear cells were cultured in suitable medium, the adherent cells exhibited either an osteoclast- or mesenchymal-like phenotype cells produced with the MSCs,which displayed a fibroblast-like morphology and expressed several MSCs-related antigens CD29,CD44,CD105, but not CD34,CD45,CD106, HLA-DR. The cells were identical to human bone marrow-derived MSCs. Conclusion MSCs in HUCB can be cultured and expanded in vitro, and could be regarded as an alternative source of MSCs for experimental and clinical applications.

关 键 词：间充质干细胞 体外培养 生物学鉴定 血源性 淋巴细胞分离液 单个核细胞 MSCs 流式细胞技术 HLA-DR 生物学特性 间充质细胞 CD105 CD106 干细胞来源 临床的应用 初步鉴定 人脐带血 表面抗原 生长曲线 细胞贴壁 细胞形态 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R329-33[医药卫生—基础医学]