骨髓间充质干细胞诱导成类髓核细胞体外的研究

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1、1万方数据目录中文摘要·······················································3Abstract······················································4英文缩略词表·······················································5前言······················································6骨髓间充质干细

2、胞诱导成类髓核细胞体外研究··························8引言······························································8材料与方法························································8结果······························································19讨论·····················

3、·········································29结论······························································35参考文献··························································36致谢······························································40综述···············

4、···············································412万方数据骨髓间充质干细胞诱导成类髓核细胞的体外研究中文摘要【目的】1.分离、培养扩增和鉴定兔骨髓间充质干细胞（BMSCs）；2.通过体外诱导BMSCs成类髓核细胞，寻找细胞体内移植的合适时间点；【方法】应用单纯贴壁培养法分离、培养、扩增兔BMSCs和髓核细胞，并应用免疫细胞化学方法检测BMSCs表面相对特异性分子(CD90、CD44、CD45、CD34)的表达率以鉴定BMSCs；将接种有BMSCs的Trans

5、well小室置于含有髓核细胞的培养皿中，建立共培养模型；制定诱导BMSCs生长曲线，用RT-PCR（reversetranscriptionPCR）和Westernblot检测其表达蛋白聚糖和Ⅱ型胶原，结合以上检测寻找最佳共培养时间，为椎间盘组织工程种子细胞的更深入研究提供实验数据。【结果】应用贴壁培养法成功分离、培养、扩增兔BMSCs和髓核细胞，免疫细胞化学检测BMSCs表面相对特异性分子的阳性表达率显示：CD90为91.2%，CD44为93.5%，CD45为4.4%，CD34为5.5%。在共培养模型中，

6、经细胞生长曲线检测、RT-PCR和Westernblot检测结果提示BMSCs与髓核细胞共培养可诱导BMSCs表达蛋白聚糖和Ⅱ型胶原，在三代以内经诱导细胞生长曲线无明显改变，结合以上检测认为细胞共培养15天左右达到最佳状态，适合进行体内移植。【结论】1.采用单纯贴壁法可以成功分离出纯度较高的BMSCs；2.在Transwell小室和培养皿构成共培养系统中，BMSCs和髓核细胞共培养可以诱导BMSCs向类髓核细胞分化；BMSCs和髓核细胞体外共培养15天左右，此时适宜作组织工程种子细胞进行体内移植。【关键词】

7、骨髓间充质干细胞髓核细胞类髓核细胞共培养3万方数据BMSCsinducedintonucleuspulposus-likecellsinvitrostudiesAbstract[Objective]1.Isolation,expansionandidentificationofRabbitBMSCs；2.BMSCswasinducedintonucleuspulposus-likecells，thenfindtherighttimeforthecelltransplantation.[Methods]App

8、licationpurestickwallculturemethodisolation,expansionrabbitBMSCsandnucleuspulposuscells，anduseimmunocytochemistryidentifyBMSCssurfacerelativespecificmolecular(CD90、CD44、CD45、CD34)todeterminethepurificationofB