I am also growing MCF10A cells. I have few questions.
1. I do have DMEM media, don't have DMEM/F12..can I just grow this cell in DMEM?
2. How should I dilute the Cholera toxin and Hydrocortisone?
3. What is the doubling time of MCF10A cells?
1. I Have not tried to grow these cells in DMEM. I suspect it would work as long as you add the other media components that are required for these cells to grow.
2. I rehydrate both the solid hydrocortisone and the cholera toxin in sterile water.
I buy 100mg solid hydrocortisone and add 2mL sterile water. I place this on a roller to mix until it is fully rehydrated. Then, in a tissue culture hood, I use a 5mL syringe and a 0.2uM filter tip to filter the redissolved hydrocortisone. I make 1mL aliqouts (the amount to add to 500mL media) in eppendorf tubes and freeze these at -20C.
I buy 1mg solid cholera toxin and add 1mL sterile water and mix it well. This is stored at +4C.
3. In my experience the doubling time of MCF10A cells is 24 to 32 hours, with later passages of these cells growing at the slower end of that range.
Edited by lynnb, 17 July 2009 - 09:48 PM.