Hello everyone,
I am trying to grow the 4T1 mammospheres in a 6 well low attachment plate.
Does serum have a negative effect on mammosphere growth?
Why the mammosphere medium DMEM/HAM F12 must be a serum free medium suplemented with EGF and bFGF?
Thanks in advance.
Cancer Stem Cells (CSCs) tend to differentiate under the influence of serum. Hence in order to avoid that, you use a serum free media and provide only the growth factors that would allow the enrichment of these stem cells. EGF and FGF in addition to Insulin are the growth factor supplements that are required for CSCs to proliferate. You could also add lif1 to the cocktail to prevent any minute chances of differentiation.
We have found that the addition of B27 supplement (Invitrogen) seems to boost sphere growth as serum would, without influencing differentiation or causing cells to stick. For mammospheres, B27 without Vitamin A is the one to go for.
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