Freezing Down Cells

(C Bennett, 1/01)

• 1. Prepare appropriate volume (1 mL/plate) of media (10% CS or 10% FCS) containing 10% DMSO and place on ice

  2. Label cryogenic vials (cell line, date and box number)

  3. Trypsinize each plate for 5 minutes

  4. Add 1 mL concentrated CS or FCS

  5. Combine all the plates and spin in 12 mL tubes with snap cap for 5 minutes at 1k rpm

  6. Aspirate media and add 1 mL 10%DMSO media/plate to cell pellet. Resuspend by gentle triteration.

  7. Aliquot 1 mL of cells per cryogenic vial

  8. Place cells at 80 C for 24-48 hours

  9. Transfer vials to liquid nitrogen tank