I am working on generating breast cancer cell lines from breast cancer tissues obtained from surgery. The tissue needs to undergo some form of disaggregation to generate single cells. These are then cultured in serum-free medium with growth factors(insulin, EGF and bFGF). The protocols I have seen used DMEM-F12 or medium199 to culture cells after the enzymatic disaggregation using collagenase/hyaluronidase. Are these the only media that can support the cell growth or other media can be used?
I would suggest to try variety of mediums to test which is the best suitable for your primary cell culture it is rather simple experiment prepare individual dishes with all the media you want to test and seed the cells into the media and see their attachment and growth after 24-48 hours. For most cells I could easily substitute the DMEM and F12 as they are very similar is composition. F12 contains Ca however so if your cells doesn't have specific calcium signalling they will most probably behave the same. Immune cells however have a tendency to activate in presence of Ca+ ions so DMEM normal glucose is usually best option for them. RPMI is more different in composition but also shuld be suitable for the majority of cancer cell lines. The word of caution though if you have any previous data say in F12 the switch to a different medium may significantly affect your experimental results as cells behave different and their mRNA expression profiles are different in different mediums.
Dear Claudia,
I think the DMEM medium is best suited to the needs of your experiments, including carbohydrate and electrolyte levels. However, overexpression of EGFR is a fairly common mutational event in breast cancer cells. Therefore, it is important to prevent depletion of the cognate ligand(s) during growth in starved cells after disaggregation. I suggest providing higher concentrations of EGF (200 micrograms/ml) in a serum-free medium. The amount of glutamine to be added can be also important.
I wish you good results for the New Year!
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