利用人骨髓间充质干细胞在生物反应器中构建小口径血管

背景:目前组织工程生物反应器已对血管、肌腱、软骨、骨、心肌瓣膜、气管、膀胱和干细胞等进行了大量的研究.目的:采用改进的生物反应器系统,应用人骨髓间充质干细胞(hBMSCs)构建小口径的组织工程血管.设计、时间及地点:单一样本观察,于2005-06/2008-03在华东理工大学机械与动力工程学院和上海组织工程研究与开发中心完成.材料:血管生物反应器为华东理工大学自制;骨髓基质干细胞来自健康志愿者的骨髓.方法:设计一套血管生物反应器系统,采用有限元方法对组织工程小血管托架材料进行分析,从而设计一套用于构建直径为2 mm的小血管托架;收集人的原代骨髓基质干细胞进行体外扩增和培养,选用第3代细胞与聚羟基乙酸酯(PGA)复合后置于血管生物反应器中动态培养;培养4周后,对材料复合物取材.主要观察指标:细胞材料复合物生长状况及hBMSCs-PGA复合物的其他相关榆测.结果:hBMSCs-PGA材料复合物培养4周取材进行大体观察和扫描电镜观察,形成血管样组织,色泽明亮,有一定的弹性,用镊子反复压下血管能够反弹恢复原样;细胞分泌的胶原基质排列较规则,免疫组织化学结果表叫血管含有平滑肌弹性肌动蛋白的成分.结论:改进的血管生物反应器能模拟血管的力学环境,并能利用hBMSCs成功构建组织工程化小血管组织.

Construction of small-sized blood vessels in a bioreactor using human bone marrow stromal stem cells

BACKGROUND: There are plentful studies about bioreactor of tissue engineering of blood vessel, tendon, cartilage, heart valve,trachea, bladder and stern cell. OBJECTIVE: To construct small-sized tissue-engineerad blood vessels with human bone marrow stromal stem cells (hBMSCs) in improved bioreactor system.DESIGN, TIME AND SETTING: The single sample observation stuay was performed at the School of Mechanical and Power Engineering, East China University of Science&Technology, and Shanghai Tissue Engineering Research & Development Center from June 2005 to March 2008.MATERIALS: Vessel bioreactor was self-made by East China University of Science&Technology. hBMSCs were harvested from healthy volunteers. METHODS: A set of support bracket constructing tissue engineered blood vessels with the diameter of 2 mm was designed with the application of Finite Element Methods as an analysis method analyzing support bracket of tissue engineered small-sized blood vessel. Primary hBMSCs were first Collected and further cultivated exvivo. The third passage cultured cells were then seeded on the polyglycolic acid (PGA) to fabricate the cell-scaffold composite. Subsequently, this composite was subjected to dynamical culture in the blood vessel bioreactor. After cultured for 4 weeks, the composite was removed from the bioreactor.MAIN OUTCOME MEASURES: The following mentioned references were measured: composite growth; other correlation detection of the hBMSCs-PGA composite, RESULTS: Gross observation and scanning electron microscope were used at4 weeks after hBMSCs-PGA composite culture. It was observed that the tissue-engineered blood vessel had a bright color and certain elasticity. The blood vessel could rebound to its odginal shape after repeated press by the forceps.The secreted collagen matrix arrayed orderly around the cells and smooth muscle elastic acttn could also be detected in the formed tissues using immunohistochemistry. CONCLUSION: The in vivo mechanics conditions of blood vessels can be simulated using the current blood vessel bioreactor system. Using hBMSCs, the construction of tissue engineered small-sized blood vessels can be successfully achieved.