Failure in cell detachment after trypsinization!
I use trypsin to harvest my cells (4T1), but they don't detach after trypsin incubation. I usually use 5ml of trypsin per 75cm2 T flask and incubate for 7 mins at 37C. Has anyone encountered the same problem? How did you solve it?
Kind Regards
Hi
I would wash the cells with PBS/EDTA previouly, if this is not working,you could place the flask for several minutes at -20°C, this worked for my cells. you should also increase the incubation time,
best regards
I usually encounter the same condition as you had especially for my PC9 cells. I think it depends on the cells and the confluency of the cells itself and the trypsin-EDTA. So, my suggestion is you should avoid warming your trypsin-EDTA for several times, because as we know, trypsin is such kind of enzyme which can be worse by heating. And also you have to incubate the cells after trypsinization is at least 10 minutes long, and after incubation, please briefly knock the bottom of the plate in order to help cell detached from the plate/flask. Hope it helps! good luck!
I know it has been a while since this question was made but I would like to update here by saying that trypsin has never worked for me and it usually made my 4T1 culture die, so I have started using a scrapper instead and it has functioned perfectly for me.
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