Passaging A1 cell

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