Hi
Osteogenic differentiation media is usually composed of B-glycero-phopshate, ascorbic acid and dexamethasone. Why is calcium not used to promote hydroxyapatite formation in vitro ?
The calcium is in the basal medium, not a separate additive like ascorbate.
Hi Ellen
Both calcium and phosphate are in the basal medium ,yet only B-glycerol phosphate is added. Is there a reason for increasing it to 10mM like some protocols suggest?
I think people still debate the amount of B-glycerophosphate to add. Without any matrix is deposited, but does not mineralize. At high concentrations, like 10 mM there is increased mineral deposition, but it may not be bonelike. Page 38 of this article has a thoughtful discussion. Article Rat Osteoblast Cultures
The cell source, culture substratum and all the additives probably have to be considered. I think the higher concentrations are used because total mineralization is higher.
Hi Greg,
I think as an ion, calcium will trigger more cell surface receptors and therefore manipulate gene expression more so than phosphate. So it has been more favourable to vary phosphate levels.
Though it would be interesting to vary the phosphate and calcium to see how this effects mineral deposition. No doubt species, batch and cell variability would also be a factor!
Good luck!
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