Thawing frozen A1 cells

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Revision as of 07:44, 24 September 2014 by Stephanh (talk | contribs)
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Materials:

  • Gelatin (warmed to 37ºC)
  • HS for A1 cells
  • 1x 15 ml FALCON tubes
  • Pipettes
  • 1x 175 cm2 flasks
  • frozen cells into liquid nitrogen

Procedure:

  • put 10 ml Gelatin into the new 175 cm2 flasks (remove the remaining liquid)
  • pipette 5 ml fresh media into the 15 ml FALCON tube
  • thaw the cells into a baker contains warm (nearly 25ºC) water until there is a very small piece of ice left
  • transfer the cells into the FALCON tube
  • centrifuge at 1000 rpm, 3 min at 4ºC
  • aspirate the supernatant
  • dissolve the pellet into 1 ml fresh media
  • fill in the new flasks 25 ml HS for A1 cells
  • transfer the cells in the flask
  • label the flask with the name of the cells, the passage number and the date
  • check the cells the next day, that they have attached on the flask

The cells grow at 25ºC, 2% CO2