软骨发生
阿格里坎
运行x2
软骨内骨化
细胞生物学
化学
肌肉肥大
硫氧化物9
细胞分化
细胞培养
II型胶原
软骨
内分泌学
内科学
基因表达
男科
细胞
解剖
生物
生物化学
基因
病理
医学
骨关节炎
遗传学
替代医学
关节软骨
作者
Faiza Altaf,Thomas M. Hering,Najam Kazmi,Jung U. Yoo,Brian Johnstone
摘要
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: Col10a1, 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.
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