Thawing cells is simple. However, speed is key to the optimum cell recovery.
- Vial of frozen cells
- 37°C Water Bath
- Tissue Culture Medium Containing 5–10% Serum or Other Protein
- Place vial of cells in 37°C water bath and agitate until thawed. It is important to thaw the cells quickly. Do NOT allow thawed cells to remain in freezing media any longer than necessary.
- Slowly add thawed cells to 9 mL of medium containing serum. Invert tube 2 or 3 times to mix, or mix gently by pipetting up and down several times.
- Centrifuge for 10 minutes at 200 x g.
- Aspirate or decant the supernatant and gently resuspend the cell pellet in 10 mL of medium.
- Remove an aliquot for cell count and proceed with experimental manipulations.
- The cell suspension may form clumps after standing at room temperature. This can be avoided by preparing and using the cells promptly, or by adding DNase to the suspension at a final concentration of 10 units per mL.