Cell Cryopreservation Medium (DMSO-free)

For the cryopreservation of cells such as PBMCs, stem cells and T cells

NMPA (National Medical Products Administration) Pharmaceutical Excipient Filing
Filing Number: F20240000500

Identity of Cell Cryopreservation Medium (DMSO-free)

Appearance	Colorless, pale yellow or yellow transparent liquid
Sterility	Sterile
Endotoxin Level	<0.125EU/mL
pH	7.2±0.5

Cell Cryopreservation Medium (DMSO-free) — A safer alternative to DMSO cell culture freezing for PBMCs, stem cells, and T cells.

Cryopreservation Protocol for Cell Cryopreservation Medium (DMSO-free)

Collect the cells to be frozen,centrifuge the cell (400×g,5 minutes,RT) and remove the supernatant.
Resuspend cell pellet with proper solution (saline,culture medium or DPBS) ,followed by cell counting and cell survival rate detection.A viability of over 90% before cryopreservation is preferred.Then centrifuge (400×g,5 minutes,RT) and remove the supernatant.
Resuspend cell pellet using the stem cell freezing media for steam cells,the recommanded cell density is 1×10⁶-1×108/mL.For PBMC and aβ-T cells,the recommanded cell density is 1×10⁶-2×107/mL.The freezing volumeshould be 0.5-1.5 mL/tube.If using a cryopreservation bag for large volume cryopreservation,please contact the technical team to obtain the programed cooling curve.Users can also add autologous plasma,serum,or HSA according to experimental needs to improve post-thaw viability.
Transfer the cryovial to a-80℃ freezer.If long-term freezing is required,transfer to liquid nitrogen after 12hours.

Thawing Protocol for Cell Cryopreservation Medium (DMSO-free)

1. After removing from liquid nitrogen,immediately immerse the cryovial into a 37℃ water bath,stir to rapidly thaw the cells within 2 minutes.

2. Transfer thawed cells to a conical tube,slowly and 9 times volume of proper diluent.

3. Equilibrate by gently invert the tube for 10 times.

4. Centrifuge (400×g,5minutes,RT) and remove the supernatant.

5. Resuspend cell pellet with proper media/solution.

6. Perform cell count test if needed.

7. Proceed for downstream application.

Viability Test for Cell Cryopreservation Medium (DMSO-free)

Add thawed cells to 4 times the volume of complete culture medium,transfer to a centrifuge tube,and centrifuge for 5 minutes.Discard the supernatant,resuspend the cells using DPBS or complete culture medium,and then test the cell survival rate.