I am using MDA-MB-231 and MCF7 for doing experiment. We have CO2 independent medium. Could you tell me how should I add other components (FBS, Glutamin/max, Penicillin, Insulin..). If I need 1ml medium, how should I dilute these components?
For FBS, Glutamin.. should I store in refrigerator (2-8 degree) for long time or I have to freeze them?
I look forward to hearing from you.
Thank you so much for your help.
Hi,
I am working with MDA MB 231 and am culturing them in a CO2 independent medium. I use Leibovitz's L15 medium. The formulation by GIBCO contains the required amount of L Glutamine and lacks sodium bicarbonate (an essential component of the bicarbonate buffer system required in CO2 dependent media).
Complete the medium with 10% FBS and 1% Pen/Strep and store at 2-8 degrees (the medium is stable for prolonged periods, you may, however, see some debris formation in the incomplete medium but that is not a point of concern). MDA MB 231 cells grow well in this medium but do not require insulin. (MDA MB 435S cells require 0.01mg/ml insulin).
You will have to culture the cells in a CO2 free incubator at 37 degrees. Simply turn off the CO2 injector or set it to 0.0.
Hope this helps.
Best wishes.
You must add the additive based on the final amount of the material in the culture medium. For example, the 1. The serum concentration at the final concentration should be 10%, it is easy to calculate the serum required by a simple mathematical procedure (10 cc for 90 cc and 50 cc for 450 cc, volume Your initial culture is 500 cc, so you need to add 5 cc to 50 cc, so you need to add 55 cc of the serum to your crop for the 500 cc medium. Note that in solutions of grams / volume, the amount of material is dissolved in 100 cc of solvent, for example 10% glucose, 10 g glugose per 100 cc solvent, but in volume / volume solutions such as 30% alcohol 30 cc of alcohol is dissolved in 70 cc distilled water. You should filter the additive material in the addition of materials to the cell culture medium; besides antibiotics .
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