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用于软骨生成的组织工程策略——使用人软骨细胞和连续细胞系的微团培养和三维培养

Tissue engineering strategies for cartilage generation--micromass and three dimensional cultures using human chondrocytes and a continuous cell line.

作者信息

Tare Rahul S, Howard Daniel, Pound Jodie C, Roach Helmtrud I, Oreffo Richard O C

机构信息

Bone and Joint Research Group, Developmental Origins of Health and Disease, University of Southampton, Southampton SO16 6YD, UK.

出版信息

Biochem Biophys Res Commun. 2005 Jul 29;333(2):609-21. doi: 10.1016/j.bbrc.2005.05.117.

Abstract

Utilizing ATDC5 murine chondrogenic cells and human articular chondrocytes, this study sought to develop facile, reproducible three-dimensional models of cartilage generation with the application of tissue engineering strategies, involving biodegradable poly(glycolic acid) scaffolds and rotating wall bioreactors, and micromass pellet cultures. Chondrogenic differentiation, assessed by histology, immunohistochemistry, and gene expression analysis, in ATDC5 and articular chondrocyte pellets was evident by the presence of distinct chondrocytes, expressing Sox-9, aggrecan, and type II collagen, in lacunae embedded in a cartilaginous matrix of type II collagen and proteoglycans. Tissue engineered explants of ATDC5 cells were reminiscent of cartilaginous structures composed of numerous chondrocytes, staining for typical chondrocytic proteins, in lacunae embedded in a matrix of type II collagen and proteoglycans. In comparison, articular chondrocyte explants exhibited areas of Sox-9, aggrecan, and type II collagen-expressing cells growing on fleece, and discrete islands of chondrocytic cells embedded in a cartilaginous matrix.

摘要

本研究利用ATDC5小鼠软骨生成细胞和人关节软骨细胞,通过应用组织工程策略,包括可生物降解的聚乙醇酸支架、旋转壁生物反应器和微团培养,试图建立简便、可重复的软骨生成三维模型。通过组织学、免疫组织化学和基因表达分析评估,在ATDC5和关节软骨细胞微团中,软骨分化明显,在嵌入II型胶原蛋白和蛋白聚糖软骨基质的腔隙中存在表达Sox-9、聚集蛋白聚糖和II型胶原蛋白的不同软骨细胞。ATDC5细胞的组织工程外植体让人联想到由许多软骨细胞组成的软骨结构,在嵌入II型胶原蛋白和蛋白聚糖基质的腔隙中,对典型软骨细胞蛋白进行染色。相比之下,关节软骨细胞外植体表现出在羊毛上生长的表达Sox-9、聚集蛋白聚糖和II型胶原蛋白的细胞区域,以及嵌入软骨基质中的离散软骨细胞岛。

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