利用脱细胞血管基质体外构建小口径组织工程血管  

Preliminary Study of in vitro Construction of Small-Diameter Vascular Graft by Seeding Cells onto Decellullarized Arterial Matrix

作　　者：武欣[1] 谷涌泉[1] 段红永[1] 陈兵[1] 李建新[1] 吴英锋[1] 张淑文[1] 汪忠镐[1] 张建[1] 

机构地区：[1]首都医科大学宣武医院血管外科,北京100053

出　　处：《中国实验动物学报》2010年第5期377-382,I0002,I0003,共8页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家高技术研究发展计划(863计划)项目(2006AA02A134);北京市自然科学基金项目(项目编号:5082007和2103049)资助

摘　　要：目的探讨利用犬的间充质干细胞诱导分化种子细胞,以异种脱细胞血管基质为基础体外构建小口径血管移植物。方法采用密度梯度离心和贴壁培养的方法从犬骨髓中分离出间充质干细胞并体外培养,诱导分化成内皮样细胞和平滑肌样细胞;采用非离子型去垢剂和胰蛋白酶去除猪颈动脉血管壁结构细胞,对脱细胞基质进行组织学、力学检测及孔隙率评估。在生物反应器内采用旋转种植的方法将犬骨髓间充质干细胞诱导的内皮样细胞种植到脱细胞基质上,体外构建小口径组织工程血管。结果犬的骨髓间充质干细胞体外能够定向诱导分化为平滑肌样细胞和内皮样细胞,可以作为血管组织工程的种子细胞。经过脱细胞处理后,光镜和电镜观察证实血管壁的细胞成分完全去除。具有良好的孔径和孔隙率。支架在生物力学、孔隙率等方面符合构建组织工程血管支架的要求。在生物反应器内剪切力条件下可以初步构建出组织工程血管。结论小口径血管移植物可以将间充质干细胞诱导种子细胞,以异种脱细胞血管支架作为基质,在搏动性生物反应器内培养的方法进行构建。Objective To explore the use of canine bone marrow mesenchyme stem cells to induce differentiated seed cells in vitro,and using the seeding cells to develop small-diameter vascular graft based on xenogenic decellularized arterial matrix.Methods Canine bone marrow mesenchymal stem cells were separated by density gradient centrifugation and cultured in vitro,and differentiated into endothelial cells and smooth muscle cells.Porcine carotid arteries were decellularized using Triton X-100 and tripsin to eliminate all the cellular components.The mechanical and biocompatibility properties of the decellularized scaffolds were identified and evaluated.Small-diameter tissue engineered vascular grafts were constructed using the decellularized scaffolds and seeding cells in the dynamic bioreactor under dynamic conditions.Results The cultured bone marrow mesenchymal stem cells were successfully differentiated into endothelial-like cells and smooth muscle-like cells,which can be used as seeding cells.Porcine carotid arteries were successfully decellularized by trypsin,ammonium hydroxide and non-ionic detergent eliminating all cellular components to make acellular scaffolds.Good pore size and porosity of the acellular scaffold were obtained.The biomechanics,immunogenicity and cell compatibility characteristics of the scaffolds could meet the requirements of tissue engineered vascular graft.Therefore,construction of the tissue engineered vascular graft in vitro in the dynamic bioreactor was succeded.Conclusions Bone marrow mesenchyme stem cells can be induced to differentiated into endothelial-like cells and smooth muscle-like cells in vitro.Using these cells as seeding cells and xenogenic decellularized arterial matrix as acellular scaffolds,small-diameter vasculargrafts can be constructed in a pulsatile bioreactor by culture under dynamic conditions in vitro.

关 键 词：间充质干细胞 脱细胞 支架 生物反应器 组织工程 

分 类 号：Q95-3[生物学—动物学] R654.3[医药卫生—外科学]