I am researching an oncosuppresive protein in skin and using HaCaT cells as the in vitro model. I am currently using normal DMEM media with 10% FBS. Should I be modifying anything in the media, especially with regard to calcium level (calcium level in DMEM is around 1.3 mM)? I have not observed any hindrance in their proliferation with current conditions.
Hi, Sutharsan,
If you are not interested in epidermal differentiation and morphogenesis, then you can safely grow the cells in your medium. Keep in mind that these cells tend to terminally differentiate in corneocytes in presence of high (i.e., > 1.5-2 mM) calcium concentration (pay attention to the fact that also the FBS contains Ca2+). However, in your conditions there should be no problems.
Good luck.
Sergio
Hi, Sutharsan,
If you are not interested in epidermal differentiation and morphogenesis, then you can safely grow the cells in your medium. Keep in mind that these cells tend to terminally differentiate in corneocytes in presence of high (i.e., > 1.5-2 mM) calcium concentration (pay attention to the fact that also the FBS contains Ca2+). However, in your conditions there should be no problems.
Good luck.
Sergio
Thanks Sergio. I have checked the Ca++ levels in our serum. 3.45mmol/l. and I add 10% FBS to the medium.
Hi, Sutharsan,
we also use HaCat cells in the lab and use normal DMEM completed with 10% FCS.
If you doubt about the Ca concentration in your FCS you can think about Ca depletion. There are existing protocols to get rid of the Ca.
Good luck
Sibylle
Hi all. does anyone knows what is the concentration of calcium in normal DMEM? (approx). i am doing calcium depletion on FBS, but i am still concern about amount of calcium in medium. thanks
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