Thawing frozen C2C12 cells: Difference between revisions
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== | == Material == | ||
* frozen C2C12 cells | * frozen C2C12 cells | ||
* HS for C2C12 cells | * HS for C2C12 cells | ||
| Line 6: | Line 6: | ||
* Cell culture flask | * Cell culture flask | ||
==Procedure | == 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 | |||
Latest revision as of 14:30, 8 June 2015
Material
- 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