Cell Preservation Medium

Cell Preparation

1.Prepare the cells to be frozen into cell suspension by mechanical or enzymatic dissociation.
2.Centrifuge the cell suspension to obtain the cell pellet.
3.Remove the supernatant. Note: Remove as much washing solution as possible to reduce the dilution of the cryopreservation medium.
Cell Cryopreservation
1.Add precooled (2 - 8°C) cryopreservation medium.
a. Cell density: The cell density should be 0.5 - 30×106 cells/mL according to the conventional cell culture protocol (it can be higher).
b. The cryopreservation medium already contains DMSO, and there is no need to add any other cryoprotectants.
2.Precooling: Incubate the cell/cryopreservation medium mixture at 2 - 8°C for about 10 minutes. 3.Programmed cooling:
a. If using a programmed cooling container, place the cryopreserved cell suspension in a pre-cooled (2 - 8°C) cooling container, and then put the programmed cooling container in an -80°C refrigerator. After 24 hours, transfer it to a liquid nitrogen tank (below -130°C) for long-term storage.
b. If using a programmed cooling device to cool the cryopreserved cell suspension at -1°C/min to -100°C, it should be immediately placed in a liquid nitrogen tank (below -130°C) for long-term storage.
c. The cryopreserved samples should be stored in a liquid nitrogen tank (below -130°C) for long-term. If placed in an -80°C refrigerator, only short-term storage (several weeks to months) is recommended.
Sample Thawing
1.Thawing and resuscitation: Quickly thaw the sample in a 37°C water bath or a similar mechanical thawing device.
a. Gently rotate the sample during thawing until all visible ice has melted. The thawing time for a 1 mL