Protocol for Recovery of Frozen Cultured Cells

1. Remove cells from liquid nitrogen
2. Thaw the vial by continuously swirling in a 37^oC water bath until on a slight amount of ice remains
3. Clean outside of tube with alcohol before opening
4. Add thawed cells to ~20ml of chilled fresh medium
5. Mix and then centrifuge (to remove freezing medium)
6. Suspend the cells in 10ml of chilled fresh medium and place in a 25 cm² flask
7. Incubate overnight using standard conditions before cell count and viability assay