Thawing frozen C2C12 cells

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  • frozen C2C12 cells
  • HS for C2C12 cells
  • pipettes and tips
  • FALCON tube 15 ml
  • Cell culture flask


  1. heat the HS up to 37ºC
  2. pipette 14 ml C2C12 HS media in a 75 cm2 cell culture flask
  3. pipette carefully 1 ml from the warm media in the cryo tube to the frozen cells
  4. pipette carefully up and down till the cells are thaw
  5. pipette all in the FALCON tube
  6. centrifuge at 1000 xg, 3 min at 4ºC
  7. label the cell culture flask with the name, date and C2C12 with the passage number
  8. aspirate of the media and resuspend the cells in 1 ml of the new HS
  9. pipette the suspension in the cell culture flask
  10. check the cells under the microscope
  11. put the flask at 37ºC, 10% CO2