同种异体软骨细胞及自体骨髓基质干细胞混合共培养构建软骨皮下移植可行性探究.doc

同种异体软骨细胞及自体骨髓基质干细胞混合共培养构建软骨皮下移植可行性探究[摘要]背景：软骨组织工程的种子细胞问题是目前研究的热点和难点，如何找到一种既能够避免对自体软骨进行取材又能够达到稳定软骨构建目的的方法呢？本研究尝试利用少量同种异体羊软骨细胞作为软骨诱导微环境提供者，与扩增后的羊自体BMSC混合共培养并植入皮下环境，探讨利用同种异体软骨细胞共培养构建软骨皮下移植的可行性。方法：本实验对山羊软骨细胞和BMSC分别进行取材和分离培养扩增，并将以上细胞分为以下四组进行混合并接种在PGA支架材料上：A组：100%自体软骨细胞；B组：30%自体软骨细胞+70%自体BMSCs；C组：30%同种异体软骨细胞+70%自体BMSCs； D组：100%同种异体软骨细胞。经过体外构建6周后植入羊皮下进行体内构建12周，对所形成的组织块进行大体观察和组织学染色等评价。结果：自体软骨细胞组和自体软骨细胞混合自体BMSC组皮下移植后可见成熟软骨组织形成，但同种异体软骨细胞参与的两组（包括同种异体软骨细胞混合自体BMSC的实验组和单纯异体软骨细胞组）在皮下环境中都因为较强的免疫反应未能形成软骨组织。结论：同种异体软骨细胞以及PGA支架材料的存在对于组织工程软骨在羊皮下环境的构建有负面影响。 [关键词]同种异体；软骨细胞；骨髓基质干细胞；软骨；组织工程 [中图分类号]Q813.1[文献标识码]A[文章编号]1008-6455（2011）01-0073-04 Feasibility study on co-culture of autologous chondrocyte with autologous BMSC to construct tissue engineered cartilage under subcutaneous environment LI Qiong,ZHANG Lu,ZHOU Guang-dong,LIU Yu,LIU Wei,CAO Yi-lin （Department of Plastic and Reconstructive Surgery,Shanghai Key Laboratory of Tissue Engineering, Shanghai 9th People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China) Abstract:BackgroundThe seed cells for cartilage tissue engineering remains a difficult problem, and how to find a way to avoid donor site morbidity by autologous cartilage harvest and steady chondrogenesis? In this study, we used a small amount of allogenic chondrocytes as a micro-environment provider of cartilage, and autologous BMSC as a guarantee for cell quantity to investigate the feasibility using allogenic chondrocytes for autologous cartilage tissue engineering.MethodsThe goat chondrocytes and BMSC were isolated, cultured and amplified. These cells were divided into the following four groups and seeded on the PGA scaffolds: Group A: 100% autologous chondrocyte; Group B: 30% autologous chondrocyte + 70% autologous BMSCs; Group C: 30% allogeneic chondrocyte + 70% autologous BMSCs; Group D: 100% allogenic chondrocytes. After in vitro culture for 6 weeks and subcutaneous implantation for 12 weeks, the constructs were harvested f