I have rat adipose-derived mesenchymal stem cells and the cells that are treated with maintenance media produced more alkaline phosphatase and stained more with Alizarin red than my ASCs treated with osteogenic differentiation media?
Maybe it happens because osteogenic markers shows the genes that has more activity in the start of differentiation process...and your osteogenic cells doesn't need a high protein production to osteogenesis as your non-osteogenic needs...Just wondering about that.
Mesenchymal stem cells tend to have a biphasic spectrum of responsiveness to inductive factors, including osteogenic factors. Therefore, if your particular preparation of MSCs has a tendency to behave in an osteogenic manner under baseline conditions, inductive factors like dexamethasone could actually inhibit this osteogenic process. Osteogenic tendencies can be gradually induced by several factors inclusing certain FBS preparations, and sustained exposure to high cell density during passage. For rat MSCs especially, culture at very low density - a good rule of thumb is have them touch no more than two other cells at the point of harvest. - this is an oldie but a goodie: Javazon et al. PMID: 11359947
without knowing the components and concentration it is difficult to say. a good starting point is dex concentration as carl said.also during expansion again as Carl eluded too, never leave the the culture be more than 60-70% confluent.
Kari Hanson, Few mesenchymal stem cells (esp. bone marrow-derived) have been reported to display a higher preponderance towards osteogenic lineage. Just as Carl Gregory mentioned, higher confluence may have influenced your cultures and additional factors cause negative effects on differentiation.
https://pubmed.ncbi.nlm.nih.gov/30779478/ this paper may be of help, they compare BMC vs MSC cultured in osteogenic media with and without dexamethasone.
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