Passaging A1 cell

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Revision as of 09:27, 25 March 2015 by Stephanh (talk | contribs)
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Materials

  • APBS (100 ml PBS + 25 ml ddwater)
  • TE in APBS (36 ml APBS + 4 ml TE)
  • Gelatin (warmed to 37ºC)
  • HS for A1 cells
  • 1x 15 ml FALCON tubes
  • Pipettes
  • 3x 175 cm2 flasks

Procedure

  • put 10 ml Gelatin into the new 175 cm2 flasks ( transfer the gelatine from the first to the second and from the second into the third → remove the remaining liquid)
  • aspirate the old media from the cells
  • wash the cells 1 times with 5 ml APBS
  • remove the APBS
  • add 5 ml TE
  • wait till the cells are detached
  • stop the TE with 5 ml HS – media
  • centrifuge at 1000 rpm, 3 min at 4ºC
  • aspirate the supernatant
  • dissolve the pellet into 3 ml fresh media
  • fill in the new flasks 25 ml HS for A1 cells
  • add into each flask 1 ml of the cell suspention

The cells grow at 25ºC