Thawing frozen A1 cells

Materials:

- Gelatin (warmed to 37ºC) - HS for A1 cells - 1x 15 ml FALCON tubes - Pipettes - 1x 175 cm2 flasks - frozen cells into liquid nitrogen

Procedure:

- put 10 ml Gelatin into the new 175 cm2 flasks (remove the remaining liquid) - pipette 5 ml fresh media into the 15 ml FALCON tube - thaw the cells into a baker contains warm (nearly 25ºC) water until there is a very small piece of ice left - transfer the cells into the FALCON tube - centrifuge at 1000 rpm, 3 min at 4ºC - aspirate the supernatant - dissolve the pellet into 1 ml fresh media - fill in the new flasks 25 ml HS for A1 cells - transfer the cells in the flask - label the flask with the name of the cells, the passage number and the date - check the cells the next day, that they have attached on the flask

The cells grow at 25ºC, 2% CO2