Dear Colleagues,
I am working with glioma cancer stem cells cultured in suspension as the spheroids. Generally, I used to prepare fresh, complete culture medium just before using to the cells - I used to supplement the mix of DMEM/F-12 GlutaMAX/B27/antibiotics with the growth factors FGF and EGF just before adding to the spheroid culture. However, I know some people make a complete medium every few weeks by adding everything (B27, antibiotics, FGF, EGF) to a newly opened bottle of medium and use this complete medium for a few weeks or so. So, should the growth factors FGF and EGF be added to the medium just before using to the cells? Or is it OK to add them every few weeks and use the complete medium for a long time?
Thank you in advance.
I used to add these critical factors just before use of medium. I store medium only for 2/3/4 days for replacing media. I do not store complete medium for weeks
I aliquot growth factors in 0.5ml tubes, frozen at -70C. I take one vial and mix with 100 ml or 200 ml of media to be used (needed in short term) within 1-3 days.
Hello Sylwia Katarzyna Król
The shelf life of growth media changes after the addition of supplements. Complete media containing protein supplements tend to degrade faster than basal media alone. So, I suggest that you add growth factors like FGF and EGF to the media just before use.
Best.
Subhash C. Juneja, Malcolm Nobre Many thanks for your answers.
Actually, I think you can add all the growth factors into your complete medium and store at 4 ℃. But the first thing you must take notice is that you should check the storage condition (the product data sheet) of all the growth factors. As far as I know, when storing at 4 ℃ refrigerator, EGF, FGF and PDGFRA are all OK before you running out the culture medium.
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