Changes

#At near confluence wash cells twice with PBS -/- and trypsinize normally.#Collect all the cells in a 15 mL falcon tube.#Centrifuge 5 min at 1500RPM to pellet cells.#Aspirate media.#Add media (1.8 mL per original plate).#Add sterile DMSO to a final concentration of 10% (0.2 mL per original plate).#Gently resuspend cells and aliquot 1 mL of suspension into cryopreservation vials.#Label vials with name, date, cell type and passage (if known).#Ensure isopropanol is added to the container to above the indicated line.#Place container with vials at -80 for 1-3 days.#Remove cells from container and place in liquid nitrogen storage.