Retroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes
Retroviral transduction with SOX9 enhances re-expression of the chondrocyte phenotype in passaged osteoarthritic human articular chondrocytes
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DOI：
10.1016/j.joca.2004.10.011
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发表时间：
2005-01-01
影响因子：
7
通讯作者：
Hardingham, TE
中科院分区：
文献类型：
--
作者：
Tew, SR;Li, Y;Hardingham, TE
目的：关节软骨细胞在单层培养中增殖，但转录因子 SOX9 的表达下降，细胞重组软骨组织的能力下降。我们研究了来自骨关节炎 (OA) 关节的广泛传代的人关节软骨细胞中的逆转录病毒 SOX9 表达是否使细胞能够在沉淀培养中重新获得软骨基质形成表型。设计：用 SOX9 逆转录病毒转导来自正常和 OA 关节的软骨细胞，并生长至第 7-10 代，然后作为 500,000 个细胞的沉淀培养 14 天。通过实时聚合酶链反应、组织学、免疫组织化学和 1,9-二甲基亚甲蓝测定法对沉淀进行分析。结果：当用 SOX9 转导并用 10% 血清培养成沉淀时，来自 OA 关节的软骨细胞显示出较高的 COL2A1 基因表达，但糖胺聚糖 (GAG) 合成较低。添加转化生长因子 β-3 和胰岛素样生长因子-1 增加了这些 SOX9 转导细胞沉淀中胶原蛋白 II 的表达和 GAG 的合成。与对照绿色荧光蛋白转导的细胞沉淀相比，细胞呈现圆形形态，并且胶原蛋白II蛋白的沉积增加。来自 OA 或健康软骨的转导软骨细胞也观察到类似的结果。 SOX9 转导的人真皮成纤维细胞没有表现出任何软骨形成反应。讨论：SOX9 转导使传代的关节软骨细胞在沉淀培养物中恢复软骨细胞表型，并形成软骨基质，通过生长因子增强软骨基质。转导后，来自 CA 关节的软骨细胞显示出与来自健康关节的软骨细胞相似的软骨形成恢复能力，这表明 CA 不会永久损害软骨细胞表型。 (C) 2004 年由 Elsevier Ltd 代表国际骨关节炎研究协会出版。
Objectives: Articular chondrocytes proliferate in monolayer culture, but the expression of the transcription factor SOX9 falls and the ability of the cells to reform cartilage tissue declines. We have investigated whether retroviral SOX9 expression in extensively passaged human articular chondrocytes from osteoarthritic (OA) joints enables the cells to regain a cartilage matrix forming phenotype in pellet culture.Design: Chondrocytes from normal and OA joints were retrovirally transduced with SOX9 and grown to passages 7-10 before being cultured as pellets of 500,000 cells for 14 days. Pellets were analysed by real time polymerase chain reaction, histology, immunohistochemistry and 1,9-dimethylmethylene blue assay.Results: Chondrocytes from OA joints displayed higher expression of COL2A1 gene when transduced with SOX9 and cultured as pellets with 10% serum, but glycosaminoglycan (GAG) synthesis was low. Addition of transforming growth factor beta-3 and insulin like growth factor-1 increased collagen II expression and GAG synthesis in these SOX9 transduced cell pellets. The cells adopted a rounded morphology and there was increased deposition of collagen II protein compared to control green fluorescent protein transduced cell pellets. Similar results were seen with transduced chondrocytes from OA or healthy cartilage. SOX9 transduced human dermal fibroblasts did not show any chondrogenic response.Discussion: Transduction with SOX9 primed the passaged articular chondrocytes to regain a chondrocytic phenotype in pellet culture and to form a cartilaginous matrix, which was enhanced by growth factors. Following transduction, chondrocytes from CA joints showed a similar capacity for chondrogenic recovery as those from healthy joints, which suggested that CA does not permanently compromise the chondrocyte phenotype. (C) 2004 Published by Elsevier Ltd on behalf of OsteoArthritis Research Society International.