CELLBANKER® 2

A. Freezing

For optimum results, cells for cryopreservation should be in log phase of growth. Similar or standard freezing protocols may be substituted.

1. Examine and make sure the cell culture free of contamination, in healthy situation and proper confluency, etc.
2. Perform a cell count to determine the viability of cells.
3. Gently pellet the cells centrifugation (3 - 5 minutes at 1,000~2,000rpm, 4℃). Remove the supernatant using a aspirator.
4. Gently suspend the cells with CELLBANKER^® cryopreservation medium (1 ml for 5x10⁵ - 5x10⁶ cells).
5. Dispense the cell suspension in 1ml aliquots to cryopreservation vials that have been labeled with the cell line name, cell concentration, passage date and other essential information.
6. Place the vials directly in a -80℃ for storage. If necessary, transfer the frozen vials to a liquid nitrogen storage tank after the vials have been frozen for at least 24 hours.

IMPORTANT: Optimum protocol may change with the cell types.

B. Thawing

1. Remove the frozen cell from storage and quickly thaw in a 37℃ shaking waterbath.
2. Immediately dilute and gently mix each 1ml of cells with 10ml of complete cell culture medium.
3. Gently pellet the cells centrifugation (3-5 minutes at 1,000 - 2,000rpm, 4℃). Remove the supernatant aspirator.
4. Gently suspend the cells with appropriate volume of complete cell culture medium. And plate in a culture flask.
5. Continue the further culture procedures according to standard protocols.