Freezing tissue culture cells

  • The day before freezing, split cells 1:10 in fresh medium (to get them in a dividing state).
  • Spin the cells down, and remove as much supernatant as possible.
  • Resuspend cells in 8% DMSO, 92% feral bovine serum at 4°C.
  • Transfer cells to tubes (0.5 ml/tube) and place in a freezer rack at -80°C.
  • Transfer the tubes to liquid nitrogen the next day.

General note: prolonged exposure to DMSO is toxic, so try to minimize the number of tubes handled at one time.