FAOGROW.SAM

GROWING FAO CELLS

FAO cells grow in RPMI-1640 media supplemented with 10% FBS (Fetal Bovine Serum).

Thaw cells in 37^oC water bath. Put the thawed cells into RPMI-1640 media + FBS. It may take several days for the cells to become confluent.

Splitting confluent cells (when maintaining a 75cm² flask of cells):

1. Aspirate off the media.

2. Wash with 5 ml PBS-CMF (sterile). Aspirate off the PBS.

3. Add 1ml trypsin and let incubate on cells until cells

round up and detach when flask is tapped lightly.

4. To stop trypsin reaction add 9ml RPMI-1640 + FBS.

To maintain cells, a 1:10 dilution is good.

5. Resuspend cells and remove 1 ml to new flask which contains 14ml media.

Freezing cells

Freeze cells by spinning down cells after media has been added to stop trypsin reaction. Resuspend cells in RPMI + 20% FBS + 10% DMSO and aliquot to cryovials (1ml/vial). Freeze at -20^oC for 1 hour, -70^oC overnight, then transfer to liquid nitrogen.