how to separate dead from viable cells in cultures of non-adherent cells - (Nov/28/2007 )
dear all
I have just cultured Jurkat cells and NCI-N417 cells after being stored at -80 for 1 year. since these cells grow as aggregates in suspension (non- adherent), I want to know if there is a way to separate dead from viable cells.
thanks
-yobou-
Just keep the culture going. Eventually live cells will takeover the whole population.
-genehunter-1-
A high-tech method would be to use fluoro-labelling and FACS.
Considering the potential for a big headache, I'd try the genehunter's method.
-swanny-
QUOTE (yobou @ Nov 28 2007, 03:31 AM)
dear all
I have just cultured Jurkat cells and NCI-N417 cells after being stored at -80 for 1 year. since these cells grow as aggregates in suspension (non- adherent), I want to know if there is a way to separate dead from viable cells.
thanks
I have just cultured Jurkat cells and NCI-N417 cells after being stored at -80 for 1 year. since these cells grow as aggregates in suspension (non- adherent), I want to know if there is a way to separate dead from viable cells.
thanks
A reliable quick alternative is to take the cell supernatant and let it settle over a 2-3 minute time frame. The viable cells will sediment first. You will of course lose large amounts of viable cells, but 99 % of the dead cells cells will be removed after you have aspirated off the supernatant.
Kindest regards
Rhombus
-Rhombus-