Difference between revisions of "Thawing frozen C2C12 cells"
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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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| Line 1: | Line 1: | ||
==Materials:== | ==Materials:== | ||
| − | + | * frozen C2C12 cells | |
| − | + | * HS for C2C12 cells | |
| − | + | * pipettes and tips | |
| − | + | * FALCON tube 15 ml | |
| − | + | * 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 | |
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