http://brugge.med.harvard.edu/system/files/protocols/MediaRecipesForMCF10ACells.pdf
Hi,
Yes, they need a lot actually. EGF, Hydrocortisone, cholera toxin, Horse donor serum and insulin. I put these inside DMEM/F12 mixture and they survive and grow well under these conditions.
Best,
Yunus
Hi Vandhana,
I used to culture MCF10A in DMEM:F12. In a 500 ml bottle I added 500 µl 10 mg/ml Insulin, 250 µl 1 mg/ml Hydrocortisone and 10 µl 1 mg/ml EGF. As Yunus suggested, horse serum is typically used but I used the regular FBS with no problem.
I did an experiment removing each of the growth factors and the limiting one is the hydrocortisone. When MCF10A are transformed by the incorporation of an oncogene such as HRAS12V, you can culture the cells WITHOUT the growth factors and they'll eventually start growing normally, which is a clear sign of their transformation.
Hope it helps!
Best,
Mariano
I would like to know if the cells need bovine insulin in particular or do they grow well with human insulin as well?
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