Recently, we have been encountering challenges in our laboratory in achieving satisfactory mineralization of MC3T3-E1 cell line. We would like to determine the optimal concentration of Ascorbic Acid and β-Glycerophosphate to achieve the desired outcomes. Additionally, we are unsure whether we should initially culture for 8 days with Ascorbic Acid only and then for another 8 days with β-Glycerophosphate only. We appreciate your assistance in resolving these issues and thank you in advance for your collaboration.
Kind regards,
Antonio.
Despite being called a "clonal" cell line, after multiple passages, you end up with a drift that generates a mosaic culture of cells that are different in their osteogenic differentiation potential. You could "re-clone" your line to identify clones that still have a good osteogenic potential: Get 30-40 single clones, expand, freeze, differentiate each clone with usual GP and vit C when confluent, and select/use only clone(s) that can differentiate and mineralize.
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