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

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