1. Quickly thaw frozen cells in a 37 °C water bath by gently agitating vial.

2. As soon as the ice crystals melt, wipe the vial with ethonal, and pipet gently into a culture flask containing prewarmed growth medium (at least 10 ml for 1 ml of frozen cells). Work under a sterile hood.

3. Change or add fresh media every 2-3 days.