Passaging A1 cell

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

• 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