effect of trypsinization on cell viability? - (Nov/27/2005 )

Hi,
After cultured my adherent cells under a certain condition, I checked cell viability by trypan blue staining.
When I directly dropped 0.4% trypan blue solution on the cell and waited for 15mins (after removing the medium and washing by PBS twice) there was little stained cells observed.
But, in the same culture condition, I saw many stained cells when cells were trypsinized and harvested in a suspension medium.
I am wondering if there was an effect of trypsinization on cell viability. Which observation should I rely on?

Hi
Tripsine can damage proteins of cell membrane, particulary after longer incubation, try to detach cells using only EDTA without any enzyme, and then stain and count your cells

I dont think you are supposed to let cells sit in trypan blue for more than 2-3 mins. Correct me if I am wrong.

Hi!!

How many times do you let the cells in the trypsine??

When I cultivate adherent cells, I use trypsine too for detach the cells of the flask. The cells incubate with trypsine only for 1-2 minutes in 37° C. Like this, the mortality is poor but maybe your lines is more sensitive..(I' ve seen that my EAhy are more sensitive than the Hela and both are adherent)

ciaciao