Abstract
The ATDC5 cell line exhibits a multistep of chondrogenic differentiation observed during endochondral bone formation. ATDC5 cell line is routinely induced to differentiate by exposure to insulin at high concentration thus limiting the ability to study nutrients and growth factors signal mechanism. We previously observed spontaneous induction of differentiation upon exposure to ascorbic acid containing alpha MEM medium. In this study, we first characterize the process of ATDC5 cells differentiation using ascorbic acid containing alpha MEM medium. Compared to high dose insulin, ascorbic acid induced mRNA expression of Collagen X by more than 2 folds. Ascorbic acid had no significant effect on the IGF‐1 or insulin receptors while high dose insulin downregulated both receptors respectively. Our findings suggest that we can employ ascorbic acid containing alpha MEM medium as a condition of choice for studying ATDC5 cells differentiation. We investigated the mechanism whereby ascorbic acid induced the differentiation of ATDC5 cells. Inhibitor of collagen synthesis completely blocked the effects of ascorbic acid on ATDC5 cells. When plates containing ascorbic acid‐condition chondrocyte matrix were used as substrate for prechondrocyte cells were able to differentiate in the absence of ascorbic acid an indication that collageneous matrix formation is essential for the induction of ATDC5 cells differentiation.
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