Has anyone experience with differentiated HepaRG cultures with lower Hydrocortisone levels then the recommended concentration of 50 µM? I want to decrease DMSO/Hydrocortisone levels to a minimum but i dont want to lose the benfits from differentiated HepaRG.
I use dexamethasone at 10-7 M, so I suspect that you are using a lot more than you need in your cultures. Differentiation of hepatocytes is normally maintained by keeping the cells densely packed. When primary hepatocytes are cultured densely I can maintain differentiation for about 7 days. If I overlay the cells, I can extend to about 14 days. If I use growth factors I can keep them much longer. Remember this is with primary cells. I have not used the hepatocyte cell line HepaRG yet.
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