I am culturing PC-3 cell lines. should i prepare a batch of 450ml media and 50ml serum and Store it @ 4 degree celsius. then use the media OR Prepare a 50ml complete media by aliquoting 5mL serum everytime ( 5ml serum is used by thawing a 50 mL aliquot made fro the big lot )
Technically which is the right way to use the complete media??.
Hi Ankit. It is always better if you make the complete media just prior to use. Do not freeze thaw serum every time. Rather, once you thaw it, make 10 ml aliquots of it and store some at 4 degrees and keep some frozen for later use. Use one aliquot at a time for making complete media. This will minimize the risk of contamination as well as protein denaturation due to repeated freeze thaw cycles.
I generally make 50 ml alicuot of serum and freeze them, and thaw one a time. Then I prepare complete medium in 50 ml falcon (45 dmem + 5 ml serum) for use
We usually aliquot serum by 50 ml and antibiotics by 5.5 ml, and keep them at -20C.
For complete cell culture medium we take 500 ml (the whole bottle) of comercial medium, add 50 ml thawn serum, 5.5 ml of antibiotic solution and other ingredients (if needed). Then it should be good to filter the whole medium. We keep it in the fridge, at +4C, do not aliquot. Usually we do many experiments, thus we do not use this medium for longer then few weeks.
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