Thawing frozen C2C12 cells: Difference between revisions

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(Created page with "==Materials:== - frozen C2C12 cells - HS for C2C12 cells - pipettes and tips - FALCON tube 15 ml - Cell culture flask ==Procedure:== - heat the HS up to 37ºC - pipette 14 ml...")
 
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==Materials:==
==Materials:==
- frozen C2C12 cells
* frozen C2C12 cells
- HS for C2C12 cells
* HS for C2C12 cells
- pipettes and tips
* pipettes and tips
- FALCON tube 15 ml
* FALCON tube 15 ml
- Cell culture flask
* Cell culture flask


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

Revision as of 09:33, 3 September 2014

Materials:

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

Procedure:

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