猪髓核基质成分诱导骨髓间充质干细胞分化为脊索样细胞  

Porcine nucleus pulposus matrix induces differentiation of bone marrow mesenchymal stem cells into notochord-like cells

作　　者：李德芳[1] 江增鑫 曾庆敏[1] 吴靖平[1] 谢晓诚 Li Defang;Jiang Zengxin;Zeng Qingmin;Wu Jingping;Xie Xiaocheng(Fudan University Jinshan Hospital,Shanghai 201508,China)

机构地区：[1]复旦大学附属金山医院

出　　处：《中国组织工程研究》2019年第33期5366-5371,共6页Chinese Journal of Tissue Engineering Research

基　　金：上海市金山区科委课题(2015-3-6),项目负责人:李德芳~~

摘　　要：背景:脊索细胞对椎间盘内严酷的内环境有良好的抵抗性,或许是治疗椎间盘退变性疾病合适的种子细胞来源。猪髓核终生含有大量脊索细胞,其基质内含有多种细胞因子可以促进骨髓间充质干细胞向脊索样细胞分化。目的:观察猪髓核基质对骨髓间充质干细胞分化的影响。方法:体外分离、培养、鉴定猪骨髓间充质干细胞;制备猪髓核粉末,并用配置好的DMEM完全培养基重悬粉末制备成含猪髓核组织的DMEM培养基,对骨髓间充质干细胞进行诱导3周,RT-PCR检测脊索细胞标记性基因鼠短尾突变体表型(T)、细胞角蛋白8及细胞角蛋白18的表达。该实验经复旦大学附属金山医院动物实验伦理委员会批准。结果与结论:(1)原代培养的骨髓间充质干细胞具有贴壁生长能力,在特定诱导条件下具有成骨、成脂分化能力,且能表达间充质干细胞表面标记物CD90、CD105;(2)在猪髓核基质成分诱导下骨髓间充质干细胞形态发生变化,表达鼠短尾突变体表型、细胞角蛋白8、细胞角蛋白18;(3)结果显示,猪髓核基质成分能诱导骨髓间充质干细胞分化为具有脊索细胞表型的脊索样细胞,为椎间盘退行性疾病治疗提供可靠的种子细胞。BACKGROUND: As notochord cells can develop resistance to a harsh microenvironment in the intervertebraldisc, it may be a suitable cell source for the treatment of degenerative diseases of the intervertebral disc. Porcinenucleus pulposus contains a large number of notochord cells throughout the life, and various cytokines in the cellmatrix can induce the differentiation of bone marrow mesenchymal stem cells into notochord-like cells.OBJECTIVE: To explore the effect of porcine nucleus pulposus matrix on the differentiation of bone marrowmesenchymal stem cells.METHODS: Porcine bone marrow mesenchymal stem cells were isolated, cultured and identified in vitro. Porcinenucleus pulposus powder was prepared, and suspended in the DMEM complete medium to prepare the DMEMmedium containing porcine nucleus pulposus matrix. Then, the bone marrow mesenchymal stem cells were induced in the prepared medium. After 3 weeks of induction, Notochord cell marker genes, including brachyury, cytokeratin 8, and cytokeratin18, were evaluated by RT-PCR. The study protocol was approved by the Animal Ethics Committee of Jinshan Hospital affiliated to FudanUniversity.RESULTS AND CONCLUSION: (1) Primary cultured bone marrow mesenchymal stem cells exhibited adherent growth. These cells had theability of osteogenic and adipogenic differentiation under special conditions, and expressed CD90 and CD105. (2) Under the induction ofporcine nucleus pulposus matrix, the cell morphology was changed, and the expression of brachyury, cytokeratin 8, and cytokeratin 18 wasenhanced. To conclude, porcine nucleus pulposus matrix can induce the differentiation of bone marrow mesenchymal stem cells intonotochord-like cells with notochord cell phenotype, which may provide a reliable cell source for the cell therapy of degenerative diseases ofthe intervertebral disc.

关 键 词：椎间盘退变 骨髓间充质干细胞 猪 髓核 脊索细胞 全骨髓培养法 鼠短尾突变体表型基因 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]