We are culturing primary human keratinocytes which we isolated from human skin. We found that keratinocytes differentiate in the presence of calcium. In the absence of calcium, within a week the keratinocytes will eventually differentiate and even start to look like fibroblasts. Is there an optimal concentration of calcium that we have to use in the medium?
I would recommend between 60 and 100 µM for 2D and 1mM for 3D
Have a look at:
Effects of extracellular calcium on the growth‐differentiation switch in immortalized keratinocyte HaCaT cells compared with normal human keratinocytes.
Ludovic Micallef et al.
https://doi.org/10.1111/j.1600-0625.2008.00775.x
Studies have shown that >1.25 can help stratification. 0.3-0.4 mM is the most commonly used concentration.
https://europepmc.org/article/med/6153576
Article Organotypic culture of normal, dysplastic and squamous cell ...
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