Hi everyone, I am growing NIH cells in FBS and are currently in passage 15. I want to use them to promote collagen I secretion. So, I seed them in wells and after overnight incubation replace their medium with one containing 50ug/ml ascorbic acid (according to literature). In parallel, I do the same procedure using NIH cells in NCS (the serum they are usually grown). My NIH cells fed with NCS/ascorbic are fine but the FBS/ascorbic die after switching them. Not sure why, any suggestions? This is supposed to be an established protocol. P.s.: the cells are fine before switching to ascorbic acid.
Where did you get your ascorbic acid? Is it cell culture grade?
Cells dying suggest something wrong with the ascorbic acid or the concentration.
Thank you for your answers!So ascorbic acid is suitable for cell culture. Yes, it probably has to do with ascorbic acid but the thing is that I am using the same concentration used in many publications to promote collagen I secretion. So it is quite weird. In publised literature they grow their cells in 10% CO2 whereas in our lab we use a 5%C02 incubator. Could that cause their death? I will try checking the pH after ascorbic acid addition,thank you!
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