For the first time, I am culturing the A431 cell. After trypsinization, I have noticed that almost all of the cells are not detached. For trypsinization, I used 0.5 trypsin 0.53 EDTA and kept the plate at 37 degree C for 5 minutes. Is it usual for this cell line? How can I detach the cell from plate with minimum cell damage? Thank you in advance.
In my experience, a plate of A431 cells can reach a quit high density of cells. Maybe you just need to use more trypsin or trypsinize them for longer? I only see the A431 cells staying attached to the plate, when they havn't been fully trypsinized.
I usually use TrypLe Express for A431 cells, which is "milder" for the cells and can be incubated with the cells for longer.
I would guess that 5 min might be too short. You might try just looking at the cells at intervals after adding the detachment solution. You should be able to see changes as the cells start to detach. I did not use your cell type, but I did use an autoclaved cell scraper to facilitate detachment, while trying to minimize exposure to trypsin. If you end up with a culture of floating cells, without scraping, you may have waited too long.
Do you use anything to pre-wash your monolayer before adding the trypsin? (to get rid of proteinase inhibitors).
Also I find that a gentle "smack" of the flask at the end of the 5 min helps with the detachment. If not, look into cell scrapers like the previous members suggested.
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