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

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

The cells grow at 25ºC, 2% CO2