I started culturing Calu3 cells recently and I'm wondering if it is normal for this cell line that there's quite a lot of cell death (or at least, floating cells which I assume are dead cells). Cells grow 1:4 per week in DMEM/F12 w/ Glutamax 10% FCS, 1x NEAA, pen/strep.
Hey, Leontien, we use the growth medium of alpha-MEM+10%FBS+1%P/S for the culture of Calu3 and the subculture ratio is 1:3 per 3~4 days. Is there any possibility that the cell death was increased by long trypsin time? Calu-3 is hardly to be trypsinized in less than 8 mins when they grow to confluent.
Hi, we have to trypsinize a long time to detach the cells and it hasn't changed since thawing. I've read online 20 minutes is not strange for these cells and this is what I also notice in the lab. May I ask which trypsine you use to detach them? I never grow cells too confluent, splitting at 70-80% max. I started out subculturing 1:3 but they've seem to start growing faster after a few passages. I'll set out a few subculture ratios next time i have to passage the cells.
Yeah, the trypsin-EDTA I use is 0.05%, might explain the long time needed for detachment. I'll see if increasing the percentage trypsin can reduce the time needed to trypsinize (but in the end might reach same results? longer time, lower concentration vs shorter time, but higher concentration). Thanks for your info, I'm going to try out a few things to see if I can get happier cells, even though they grow quite fast as it is now.
Hi, I seems to have the same problem with my Calu-3 cells. I used 0.05% trypsin. I am hesitating whether I should increase the concentration of the trypsin. I am afraid that it may injure the cells. May I know your opinion regarding this matter? Thank you so much.
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