Freezing of C2C12 cells

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Revision as of 08:36, 27 March 2015 by Stephanh (talk | contribs)
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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