骨髓基质干细胞扩增体系研究  

Research on the Amplification System of Bone Marrow Stromal Stem Cells

作　　者：王婷婷[1,2,3] 何爱娟[2,3] 刘豫[2,3] 曹谊林[2,3] 唐胜建[1] 周广东[1,2,3] 

机构地区：[1]潍坊医学院整形外科研究所,山东省潍坊市261042 [2]上海交通大学医学院附属第九人民医院整复外科,上海市组织工程研究重点实验室,上海市200011 [3]组织工程国家工程研究中心,上海市200241

出　　处：《组织工程与重建外科杂志》2016年第3期152-155,159,共5页Journal of Tissue Engineering and Reconstructive Surgery

摘　　要：目的比较三种不同培养体系对人骨髓基质干细胞（hBMSCs）增殖能力、表面标志和分化潜能的影响,探索hBMSCs适合的培养体系。方法获取hBMSCs,分别用DMEM、DMEM＋碱性成纤维细胞生长因子（bFGF）和无动物组份的间充质干细胞培养基（Mesenchymal Stem Cell Medium-animal component free,MSCM-acf）进行培养,反复传代至第6代,分别计数每种培养体系每个代次的细胞得率。取第4~6代细胞,用流式细胞仪对细胞表面标志进行检测,同时对各组第6代细胞分别向成软骨、成脂和成骨方向进行诱导分化。结果培养至第2代后,MSCM-acf培养体系细胞得率显著大于其他两组;流式细胞分析结果表明,MSCM-acf培养体系表面阳性标志CD73、CD90和CD105均大于99%,总的阴性标志表达小于2%,优于其他两组,尤其是DMEM＋bFGF培养体系;三组均保留了多向分化潜能,添加bFGF的培养体系成软骨分化明显优于其他两组,MSCM-acf培养体系成脂和成骨分化优于其他两组。结论 MSCM-acf培养体系可以实现hBMSCs干性维持下的大量扩增,是hBMSCs较为理想的培养体系。Objective To compare the effects of three different culture systems on the proliferation ability, surface marker and differentiation potential of bone marrow stromal stem cells（hBMSCs）, and to explore a suitable expansion system for hBMSCs. Methods hBMSCs were obtained and cultured respectively in three culture systems： DMEM, DMEM ＋basic fibroblast growth factor（bFGF） and Mesenchymal Stem Cell Medium-animal component free（MSCM-acf）, to the sixth generation of repeated passages. The cell yield of each culture system for each rate was counted. Cell surface markers of P4-P6 were detected using flow cytometry. At the same time, hBMSCs of P6 in each culture system were induced for chondrogenic differentiation, osteogenic differentiation and adipogenic differentiation. Results The cell yield in MSCM-acf group was far greater than that of the other two groups; The positive indicators of hBMSCs were detected by flow cytometry,in MSCM-acf group, CD73, CD90 and CD105 were all greater than 99%, the sum of negative cocktails was lower than 2%,which showed better results compared with the other two groups, especially DMEM＋bFGF group. All of the three groups had the potential of differentiation. Cartilage differentiation in DMEM＋bFGF was better. Adipogenic differentiation and osteogenic differentiation in MSCM-acf group were better than the other two expansion systems. Conclusion MSCM-acf expansion system can achieve a large number of hBMSCs amplification, and maintain the activity of cells. Therefore, MSCM-acf can be considered as an ideal hBMSCs expansion system.

关 键 词：骨髓基质干细胞 培养体系 细胞表面标志 分化潜能 

分 类 号：Q813.11[生物学—生物工程]