摘要
目的探讨运用组织工程技术以骨髓间充质干细胞和去细胞猪心脏瓣膜支架构建组织工程心脏瓣膜的方法。方法体外培养、扩增自体骨髓间充质干细胞并鉴定,接种于以曲拉通-脱氧胆酸钠、RNaseⅠ和DNaseⅠ去细胞的猪心脏瓣膜支架上,形成细胞材料复合物,体外培养7d,将该复合物移植于裸鼠腹腔,4周后行组织学检测。结果骨髓间充质干细胞具有与成纤维细胞相似的生物学特性,表达ASMA,Vimentin抗原,去细胞瓣膜去细胞彻底,骨髓间充质干细胞可在支架表面良好生长,形成表面光滑的细胞层,分泌细胞外基质,生长好。在裸鼠体内可继续扩增,形成多层细胞。结论运用组织工程技术以骨髓间充质干细胞和去细胞猪心脏瓣膜可生成初级组织工程瓣膜。
Purpose The objective of this study was to investigate the feasibility of creating tissue engineered heart valves from human mesenchymal stem cells and decellularized porcine heart valve leaflets to obtain viable constructs. Methods Mesenchymal stem cells were isolated, expanded and characterized by immunofluorenscence. Porcine heart valves were decellularized by Triton, RNase and DNase,reseeded and cultured under static conditions for 7 days. Then cell-scaffold constructs were implanted into peritoneal cavity above liver of nude mice for 4 weeks. Results Mesenchymal stem cells were positive＇for a-smooth muscle actin and vimentin. Histology examination demonstrated the removal of almost all cells after decellularization with preservation of the extracellular matrix. Histological examination of the tissue engineered heart valve leaflet demonstrated viable tissue and extracellular matrix formation. Scanning electron microscopy demonstrated viable secretionally active cells and confluent tissue surface. Cell-scaffold constructs retrieved 4 weeks after implantation formed three dimensional tissue. Conclusions Mesenchymal stem cells can be easily isolated,expanded and can adhere to and populate on decellularized porcine heart valve leaflet to form tissue. Generation of active tissue engineering heart valve from human mesenchymal stem cells and heart valve leaflet is feasible in in vitro environment.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2007年第2期183-186,共4页
Fudan University Journal of Medical Sciences
基金
上海市科委自然科学基金(03ZR14103)
复旦大学研究生创新基金项目(CQF152805)
关键词
骨髓间充质干细胞
去细胞瓣膜
组织工程
mesenchymal stem cells
decellularized porcine heart valve
Tissue engineering
