We have cultured the cells with 10% and 20% FBS and we've been renewing the medium once or twice a week but the cells are almost not growing at all. The medium contains DMEM + 10% FBS + 1% NEAA + 1% NaPyr + 1% P/S. Also, not all of the adhered cells seem to have acquired the reported cell line morphology. What can we do to make them grow faster?
Hello Ricardo,
We use also Caco-2 in our lab but we use only 10% FBS in DMEM with glutamine and glucose. We split our cells 1:6 every 3-4 days and use 1xTrypsin/EDTA (Sigma) for cell passage (10 min; 37°C). Stop the trypsin reaction with medium which contains FBS.
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You can check your inkubator for CO2 concentration. Try to measure the concentration in the inkubator with a hand-held measuring device. Our cells didn`t grew due to a broken CO2-sensor... The incubator showed 5% but the concentration was another...
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We don`t use heat-inactivated FBS but we test different charges. We buy different charges of FBS (100 ml) and we seed our cells in medium containing differnt charges) on 96-well-plate. Than we check with an MTT test. We buy huge amounts of the "best" FBS. So we are sure, that our FBS is not a factor which can influence the results of our experiements due to a bad growth....
Best regards,
Constanze
Not sure if this would work for Caco-2 cells as I haven't worked with them, but for the HaCaT human keratinocyte skin cells that I work with, we supplement their media with GlutaMAX (ThermoFisher) to help them grow with better stability. It has also been said that GlutaMAX enables them to grow faster, though I am not sure if that is totally accurate. However, it might be worth searching it up and seeing if anyone uses it with Caco-2 cells in the literature. All the best!
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