Hi, I am currently working with two neuronal cell lines, N2A and A1A1V. For N2A cells we are using 50% OPTIMEM + 50% DMEM +10 % FBS and 1% P/S antibiotic and for A1A1V cells we are using just DMEM with 10%FBS +1 % P/S antibiotic. Until now, the cells were growing well in these media. However, for the last two weeks, we can see cells differentiating and are overall unhealthy. Recently we got a new stock of DMEM and FBS (as we were out of the old stock) and still we hardly see any improvement. We have checked incubators for CO2 levels and temperatures (37°C for N2A cells and 33°C for A1A1V cells). We have also tried using cells from a lower passage but we still see cell death, unwanted differentiation, and clumping. Thanks for any help you can offer.
I have only experience with the N2A cell line. we have kept the, either in DMEM with 10% FBS or OPTIMEM with 5% FBS at 37°C. Your problem is most likely the new batch of FBS. FBS varies from batch to batch. I recommend to ask for 2 to 3 aliquots from your supplier before you run out of serum. For now, try to get some test aliquots and hope that your cells recover in one of them.
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