Ascorbate-enhanced chondrogenesis of ATDC5 cells
Ascorbate-enhanced chondrogenesis of ATDC5 cells
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DOI：
10.22203/ecm.v012a08
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发表时间：
2006-07-01
影响因子：
3.1
通讯作者：
Johnstone, B.
中科院分区：
文献类型：
--
作者：
Altaf, F. M.;Hering, T. M.;Johnstone, B.
ATDC5 细胞系表现出在软骨内骨形成过程中观察到的多步软骨形成分化。然而，需要长达两个月的时间才能完成细胞扩增、添加胰岛素促进分化以及进一步改变培养条件以有效诱导肥大的过程。我们试图在更实际的时间内以更简单的条件产生具有显着肥大分化的一致软骨形成。通过添加抗坏血酸，软骨形成前增殖期从 21 天缩短至 7 天，添加胰岛素后，软骨形成期会产生数量更多、尺寸更大的软骨结节。免疫组织化学表明，与对照组相比，基质精细化程度更高，sox9、聚集蛋白聚糖和 II 型胶原蛋白的 mRNA 表达均较早显着增加，且程度更高。此外，还存在强烈的肥大诱导作用：Coll0a1、Runx2 和 Mmp13 均在 7-10 天内被诱导。总之，向 ATDC5 培养物中添加抗坏血酸缩短了软骨形成前增殖阶段，产生较早的软骨形成分化，提高了基因表达和强劲的肥大分化，消除了延长培养时间和改变培养条件的需要。这种简单的修饰大大增强了该细胞系用于软骨形成研究的实用性。
The ATDC5 cell line exhibits the multistep chondrogenic differentiation observed during endochondral bone formation. However, it takes up to two months to complete the process of cell expansion, insulin addition to promote differentiation and further changes in culture conditions effectively to induce hypertrophy. We sought to produce consistent chondrogenesis with significant hypertrophic differentiation with simpler conditions in a more practical time period. By adding ascorbate, the prechondrogenic proliferation phase was shortened from 21 to 7 days, with production of cartilaginous nodules during the chondrogenic phase, after insulin addition, that were greater in number and larger in size. Immunohistochemistry indicated much greater matrix elaboration and the mRNA expression of sox9, aggrecan and collagen type II were all significantly increased earlier and to a much higher degree when compared with controls. Moreover, there was a robust induction of hypertrophy: Coll0a1, Runx2 and Mmp13 were all induced within 7-10 days. In conclusion, addition of ascorbate to ATDC5 cultures shortened the prechondrogenic proliferation phase, produced earlier chondrogenic differentiation, heightened gene expression and robust hypertrophic differentiation, abrogating the need for extended culture times and the changes in culture conditions. This simple modification considerably enhances the practicality of this cell line for studies of chondrogenesis.