Freezing Cells

1. Put freezing container on ice.

2. Make sure cells are healthy (feed 1-2 days prior to freezing).

3. Spin cells in closed conical tube @ 1200 rpm for 5 mins.

4. Label cryogenic vials (vials for freezing) with pencil under hood. Note: Loosen caps on vials for later use.

5. Put freezing medium on ice (90% FBS, 10% DMSO).

6. Pour supernatant from conical tube into waste container under the hood. Tap the vial lightly to loosen the
pellet.

7. Add freezing medium to pellet. Quickly add 1.5 ml into each freezing vial and put on ice. Leave on ice for ~
10 mins. (Note: Add enough freezing medium proportional to the amount of cell mixture for freezing).

8. Put vials in freezer container.

9. Freeze at -80° overnight.

10. Put into liquid nitrogen.

By: Olivia Granillo