Phenotypic characterization of human chondrocyte cell line C-20/A4:: A comparison between monolayer and alginate suspension culture
Phenotypic characterization of human chondrocyte cell line C-20/A4:: A comparison between monolayer and alginate suspension culture
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DOI：
10.1159/000081717
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发表时间：
2004-01-01
影响因子：
2.7
通讯作者：
Aigner, T
中科院分区：
文献类型：
--
作者：
Finger, F;Schörle, C;Aigner, T
应用DNA微阵列技术对人软骨细胞系C-20/A4的分子表型进行了研究。用Clontech人类癌细胞阵列1.2和定量聚合酶链式反应检测在单层血清和藻酸盐微球中培养的C-20/A4细胞的基因表达谱。在单层培养中，与原代培养的成人关节软骨细胞相比，参与细胞增殖的基因表达显著上调。在细胞周期调控基因中，只有两个基因，CDK调节亚基和组蛋白H4，在藻酸盐珠培养后下调，这与这些细胞在悬浮培养中增殖的能力一致。相反，当C-20/A4细胞在藻酸盐悬浮培养中时，参与细胞外基质形成的几个基因，包括基质金属蛋白酶-14、COL6A1、纤维连接蛋白、双聚糖和核心蛋白的表达上调。此外，已知在原代软骨细胞中表达的Nexin-1、Vimentin和IGFBP-3在我们的研究中也有差异表达。与该细胞系的增殖表型一致，在有血清存在的情况下，参与基质合成和周转的基因很少高表达。这些结果表明，永生化软骨细胞株，而不是替代原代软骨细胞，可以作为推广使用原代软骨细胞所不能实现的软骨细胞功能的模型。版权所有(C)2004 S.Karger AG，巴塞尔。
DNA microarray analysis was used to investigate the molecular phenotype of one of the first human chondrocyte cell lines, C-20/A4, derived from juvenile costal chondrocytes by immortalization with origin-defective simian virus 40 large T antigen. Clontech Human Cancer Arrays 1.2 and quantitative PCR were used to examine gene expression profiles of C-20/A4 cells cultured in the presence of serum in monolayer and alginate beads. In monolayer cultures, genes involved in cell proliferation were strongly upregulated compared to those expressed by human adult articular chondrocytes in primary culture. Of the cell cycle-regulated genes, only two, the CDK regulatory subunit and histone H4, were downregulated after culture in alginate beads, consistent with the ability of these cells to proliferate in suspension culture. In contrast, the expression of several genes that are involved in pericellular matrix formation, including MMP-14, COL6A1, fibronectin, biglycan and decorin, was upregulated when the C-20/A4 cells were transferred to suspension culture in alginate. Also, nexin-1, vimentin, and IGFBP-3, which are known to be expressed by primary chondrocytes, were differentially expressed in our study. Consistent with the proliferative phenotype of this cell line, few genes involved in matrix synthesis and turnover were highly expressed in the presence of serum. These results indicate that immortalized chondrocyte cell lines, rather than substituting for primary chondrocytes, may serve as models for extending findings on chondrocyte function not achievable by the use of primary chondrocytes. Copyright (C) 2004 S. Karger AG, Basel.