肌腱细胞外基质和拓扑结构诱导人脂肪干细胞成肌腱分化的实验研究

项目名称： 肌腱细胞外基质和拓扑结构诱导人脂肪干细胞成肌腱分化的实验研究

项目编号： No.31470943

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 刘伟

作者单位： 上海交通大学

项目金额： 90万元

中文摘要： 人脂肪间充质干细胞（hAT-MSC)来源丰富、扩增能力强和具有向肌腱分化的能力。我们提出科学假说：腱腱细胞产生的胞外基质（ECM)能够有效诱导hAT-MSC向肌腱细胞分化，而Biglycan/Fibromodulin是关键的诱导分子。关键ECM分子与刻纹表面结构相结合能更加有效综合模拟肌腱微环境中的理化诱导信号，促进hAT-MSC在二维模型中成肌腱定向诱导分化。利用该理念制备三维诱导支架复合力学刺激信号能精确综合模拟肌腱组织所处的体内微环境从而促进ASC在三维模型中的肌腱形成。我们拟采用CD45-/CD34+/ CD31-作为标志物和间充质细胞培养体系分选、富集和扩增出hAT-MSC作为研究对象，采用肌腱细胞产生的胞外基质脱细胞后作为EMC铺层，以及重组ECM分子铺层和siRNA沉默ECM关键分子基因等手段锁定关键的诱导分子及探讨其相关机制，并与拓扑、力学物理信号相结合验证此综合模拟假说。

中文关键词： 组织工程；脂肪干细胞；间充质干细胞；肌腱组织工程；干细胞微环境

英文摘要： Human adipose tissue derived mesenchymal stem cells (hAT-MSC) is rich in their resource, and great potential of multiple differentiation (including tenogenic differentiation) and proliferation. We propose the hypothesis: Extracellular matrix (ECM) produced by cultured tenocytes is able to induce tenogenic differentiation of hAT-MSCs and Biglycan and Fibromodulin are likely to be the key inductive molecules. Combination of key inductive molecules and microgrooved topography is able to better mimic the physical and chemical induction signals of the tendon niche environment and thus to promote tenogenic differentiation of hAT-MSCs in a 2D culture model. Further, biomimic tenogenically inductive scaffold developed based on this concept combines with mechanical loading signaling can further optimize the signal mimicking of tendon in vivo niche environment and thus promote tenogenic differentiation and tendon formation by hAT-MSCs in a in vivo setting. We plan to isolate and enrich hAT-MSC using CD45-/CD34+/ CD31- as the markers, and also use decellulized ECM produced by cultured tenocytes to serve as an inductive coating. Additionally, coating with recombinant tendon ECM molecule and the siRNA gene silence of ECM molecules will also be conducted to narrow down the key inductive ECM molecules. Moreover, combination of ECM induction with physical inductive signals of topographic structure and mechanical loading will be our ultimate strategy for comprehensive mimicking of tendon niche environment to prove our proposed hypothesis.

英文关键词： tissue engineering;adipose derived stem cells;mesenchymal stem cells;tendon engineering;stem cell niche