Reviving Cells from Liquid Nitrogen
Posted 06 August 2001 - 09:00 PM
I need some help in reviving my CHO and HEK cells after storage in liquid nitrogen in 40%serum, 10% DMSO. Any suggestions?
Posted 13 September 2001 - 09:00 PM
Posted 06 February 2002 - 02:20 PM
1) warm them up quickly - I hold them in the water bath and keep shaking. As soon as they thaw then pipette into a tube with medium immediately.
2) don't seed the cells at too low a density.
3) use a higher than normal serum concentration for the seeding (ie 15% rather than 10%)
3) it is generally recommeded that you change the medium as soon as the cells are adherant (the next day) so that the DMSO is removed.
If you are still having problems it may not be the reviving as much as it is the original freezing thats the problem.