Passaging A1 cell

From Tanaka Wiki
Jump to: navigation, search

Material

  • 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

  1. 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)
  2. aspirate the old media from the cells
  3. wash the cells 1 times with 5 ml APBS
  4. remove the APBS
  5. add 5 ml TE
  6. wait till the cells are detached
  7. stop the TE with 5 ml HS – media
  8. centrifuge at 1000 rpm, 3 min at 4ºC
  9. aspirate the supernatant
  10. dissolve the pellet into 3 ml fresh media
  11. fill in the new flasks 25 ml HS for A1 cells
  12. add into each flask 1 ml of the cell suspention

The cells grow at 25ºC