Difference between revisions of "Freezing of C2C12 cells"
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| Line 11: | Line 11: | ||
==Procedure:== | ==Procedure:== | ||
| − | + | * wash the cells with 5 ml PBS | |
| − | + | * remove the PBS and add 2 ml TE | |
| − | + | * let it incubate for 3 – 5 min | |
| − | + | * stop the TE with 5 ml HS – media and resuspend the cells | |
| − | + | * pipette the cell suspension back in the FALCON tube | |
| − | + | * centrifuge the cells (1000 xg, 3 min at 4 ºC) | |
| − | + | * remove the old media | |
| − | + | * resuspend the cells 1 ml freezing media | |
| − | + | * quickly pipette the liquid in the cryo tube | |
| − | + | * put the cells in the freezing box and store this for one day at –80 ºC | |
| − | + | * place the cells one day later into the liquid nitrogen | |
Revision as of 07:41, 24 October 2014
Materials:
- 15 ml FALCON tube
- PBS w/o Ca2+ and Mg2+
- TE in PBS
- Freezing media (10% DMSO + 40% serum + 50% DMEM)
- Pipettes and tips
- Glas pipetts
- cryo tube
- ice
- freezing box
Procedure:
- wash the cells with 5 ml PBS
- remove the PBS and add 2 ml TE
- let it incubate for 3 – 5 min
- stop the TE with 5 ml HS – media and resuspend the cells
- pipette the cell suspension back in the FALCON tube
- centrifuge the cells (1000 xg, 3 min at 4 ºC)
- remove the old media
- resuspend the cells 1 ml freezing media
- quickly pipette the liquid in the cryo tube
- put the cells in the freezing box and store this for one day at –80 ºC
- place the cells one day later into the liquid nitrogen