微载体复合间充质干细胞构建3D细胞球对体外周围神经再生的影响  

Role of 3D cell spheres constructed by microcarrier and mesenchymal stem cells on peripheral nerve regeneration in vitro

作　　者：刘平 彭江[2] 王玉[2] 许文静[2] 孟昊业[2] 杜亚楠[3] 侯昂扬[1] 常峰 LIU Ping;PENG Jiang;WANG Yu;XU Wen-jing;MENG Hao-ye;DU Ya-nan;HOU Ang-yang;CHANG Feng(Shanxi Medical University,Taiyuan 030001,China;Orthopaedic Institute,General Hospital of PLA,Beijing 100853,China;Qinghua University,Beijing 100084,China;People's Hospital Affiliated to Shanxi Medical University,Taiyuan 030012,China)

机构地区：[1]山西医科大学,030001 [2]中国人民解放军总医院骨科研究所,100853 [3]清华大学,100084 [4]山西医科大学附属人民医院,030012

出　　处：《中国矫形外科杂志》2018年第24期2277-2281,共5页Orthopedic Journal of China

基　　金：国家973项目(编号:2014CB542201);国家重点研发计划(编号:2017YFA0104702;2016YFC1101601)

摘　　要：[目的]探索微载体复合间充质干细胞构建三维(three-dimension, 3D)细胞球对周围神经再生的体外实验研究。[方法]选取48 h SD乳鼠,腹股沟处取脂肪组织,经消化、过滤、离心、分离培养得到脂肪间充质干细胞,然后将脂肪间充质干细胞与微载体构建3D细胞球,将构建好的3D细胞球与背根神经节共培养,根据添加不同培养方式的细胞球而分成3组,具体分组如下:单纯背根神经节培养组,2D细胞与背根神经节共培养组,3D细胞球与背根神经节共培养组。[结果]单纯背根神经节培养组轴突生长长度(341.66±9.01)μm, 2D细胞与背根神经节共培养组轴突生长长度(395.33±24.9)μm, 3D细胞球与背根神经节共培养组轴突生长长度(487.00±5.00)μm。3D细胞球组对背根神经节轴突生长的促进作用较2D共培养组更明显,差异有统计学意义(P<0.05)。[结论]基于微载体的3D细胞球对背根神经节轴突生长作用更明显,为以后神经组织工程治疗神经缺损提供良好的种子细胞,从而提高神经再生的能力。[Objective] To explore the role of three-dimensional(3D) cell spheres constructed by microcarrier and mesenchymal stem cells(MSCs) on peripheral nerve regeneration in vitro. [Methods] The MSCs, extracted and isolated from groin adipose tissue of new born SD rats in 48 hours, were cocultured with microcarrier to construct 3D cell spheres. In addition, the cells were isolated from dorsal root ganglion(DRG) of new born rats in 12 hours. According to the different culture conditions,the axonal regeneration of the DRG cells was compared among following 3 groups: the simple culture of DRG cells, the coculture of DRG cells with 2D MSCs, and the coculture of DRG cells with the 3D cells spheres. [Results] At 4 days of culture, the length of regenerating neurites proved(341.66±9.01) μm in the simple DRG cells group,(395.33±24.9) μm in the coculture with 2D MScs group, and(487.00±5.00) μm in the coculture with 3D cell spheres group. The effect of 3D cell spheres was significantly superior to the 2D MSCs for improving axonal regeneration of DRG cells in vitro(P<0.05). [Conclusion] The 3D cell spheres based on microcarrier has more obvious effect on the growth of neurites in the dorsal root ganglion, which might provide high quality seed cells for neural tissue engineering in treatment of nerve defects, and improve the capacity of nerve regeneration.

关 键 词：微载体 脂肪干细胞 3D细胞培养 背根神经节培养 

分 类 号：R318[医药卫生—生物医学工程]