Isolation and chondroinduction of a dermis-isolated, aggrecan-sensitive subpopulation with high chondrogenic potential
Isolation and chondroinduction of a dermis-isolated, aggrecan-sensitive subpopulation with high chondrogenic potential
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DOI：
10.1002/art.22300
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发表时间：
2007-01-01
影响因子：
--
通讯作者：
Athanasiou, Kyriacos A.
中科院分区：
文献类型：
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作者：
Deng, Ying;Hu, Jerry C.;Athanasiou, Kyriacos A.
客观的。开发一种从皮肤来源的细胞产生组织工程关节软骨结构的工艺。方法。使用改良的快速粘附过程分离真皮分离的聚集蛋白聚糖敏感（DIAS）细胞。通过定量逆转录酶-聚合酶链反应、酶联免疫吸附剂测量软骨形成潜力。测定和免疫组织化学。使用荧光显微镜检测丝状肌动蛋白（F-肌动蛋白）和纽蛋白组织。结果。快速粘附过程导致了 DIAS 细胞的选择，占整个细胞群的 10% 以下。 DIAS 细胞表现出更大的软骨诱导潜力，在聚集蛋白聚糖包被的表面上形成大量软骨细胞结节就证明了这一点。此外，与未纯化的真皮细胞相比，这些细胞在软骨细胞标记方面显示出更高的基因表达和蛋白质产量。在 DIAS 细胞和软骨细胞之间观察到类似的 F-肌动蛋白和纽蛋白组织模式。来自软骨诱导的 DIAS 细胞的三维构建体比来自其余真皮群体的构建体产生更大量的软骨基质。结论。这些发现显示了使用 DIAS 细胞共同形成组织工程关节软骨结构的一系列步骤。由于皮肤为组织工程提供了一种微创且相对丰富的细胞来源，因此这项研究提供了一种有效且稳定的技术的证据，该技术可以形成软骨结构，以便将来用来自皮肤的自体细胞进行软骨再生。
Objective. To develop a process that yields tissue-engineered articular cartilage constructs from skin-derived cells.Methods. Dermis-isolated, aggrecan-sensitive (DIAS) cells were isolated using a modified rapid adherence process. The chondrogenic potential was measured by quantitative reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent. assay, and immunohistochemistry. Filamentous actin (F-actin) and vinculin organization was detected using fluorescence microscopy.Results. The rapid adherence process led to a selection of DIAS cells, < 10% of the entire population. DIAS cells displayed greater chondroinduction potential, as evidenced by the formation of large numbers of chondrocytic nodules on aggrecan-coated surfaces. In addition, these cells showed higher gene expression and protein production in terms of chondrocytic markers when compared with unpurified dermis cells. Similar patterns of F-actin and vinculin organization were observed between DIAS cells and chondrocytes. Three-dimensional constructs from chondroinduced DIAS cells produced greater amounts of cartilage matrix than constructs from the rest of the dermis populations.Conclusion. These findings show a series of steps that work together to form tissue-engineered articular cartilage constructs using DIAS cells. Since skin presents a minimally invasive, relatively abundant cell source for tissue engineering, this study offers evidence of an efficient and stable technique to form cartilage constructs for future cartilage regeneration with autologous cells from skin.