三维立体空间动态培养及骨髓间充质干细胞软骨向分化的研究  

The effect of three - dimensional dynamic culturing of bone marrow mesenchymal stem cells on differentiation ofchondrocyte

作　　者：吕昌伟[1] 胡蕴玉[1] 崔玉明[1] 白健萍[1] 吕荣[1] 

机构地区：[1]第四军医大学骨科研究所,陕西西安710032

出　　处：《生物医学工程与临床》2008年第4期271-274,F0002,共5页Biomedical Engineering and Clinical Medicine

摘　　要：目的三维立体空间动态诱导骨髓间充质干细胞软骨向分化,分析培养状态对软骨向分化的影响。方法分离培养骨髓间充质干细胞(MSC),以海藻酸钠凝胶载体诱导软骨向分化,于转壁反应器制造周期应力性三维立体环境,比较凝胶立体结构、细胞聚集状态、细胞构型等因素对软骨向分化的影响,分别以凝胶微球悬浮高密度细胞、凝胶包埋离心细胞团、凝胶覆盖细胞及普通细胞培养等方式,进行空间动态诱导培养;同时各培养方式分别设立相应静止培养方式作为对照。10d后观察大体和组织学形态,测量生物化学指标并筛选。结果三维诱导方式比平面诱导有效,动态诱导环境优于静止诱导环境,结合三维立体空间培养和动态环境可进一步提高软骨诱导效率。其中,凝胶微球悬浮高密度细胞的培养方式分化效果最好,优于凝胶包埋细胞团的培养方式。结论海藻酸钠凝胶微球立体动态培养可有效诱导MSC的软骨向分化,为改进软骨组织工程种子细胞的立体空间动态培养提供新思路。Objective To establish suitable three-dimensional （3D） dynamic culturing of mesenchymal stem cells （MSC） for chondrocyte orientational differentiation in vitro. Methods The MSC were isolated and cultured in,and wall rotating bioreactor vessel in 3D dynamic inducing environments. Alginate gel was adopted as 3D scaffold and every parameters （such as alginate architecture,cell aggregated condition and cell construction） were analyzed. Cells were suspended and solidified in alginate droplets,imbedded in alginate mass in cell-droplet forms,covered by alginate gel on plates,or simply on plate surface. Then, cells were cultured in dynamic or static condition respectively. Gross appearance,histological examination,and biochemical measurement were observed to compare the differentiation effects. Results The 3D induction was more preferable than 2D induction,the dynamic condition was also more effective than static condition,and the dynamic 3D culturing obviously improved chondrocyte differentiation. The alginate droplets form was more useful than cell-droplet in alginate mass. Conclusion The 3D dynamic inducing could improve MSC orientation to chondrocyte differentiation and may enlightened the idea of 3D cell culturing for cartilage tissue engineering.

关 键 词：软骨细胞 分化 三维培养 间充质干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]