Composition, kit and method for cryopreserving cells
The present disclosure provides for a cryopreservation composition comprising a permeating cryoprotectant, a saccharide and a macromolecule. The cryopreservation composition has a low toxicity to cells and tissues, and promotes survival and retention of viability of the biological material during cryopreservation.
1. A cryopreservation composition comprising:
(1) about 2% (w/v) to about 20% (w/v) of polyethylene glycol;
(2) about 0.1 M to about 0.5 M of a saccharide; and
(3) about 1% (w/v) to about 10% (w/v) of gelatin,
wherein the unit of % (w/v) and the unit of M are based on the total volume of the cryopreservation composition.
2. The cryopreservation composition of claim 1 , wherein the saccharide comprises sucrose, trehalose, or a combination thereof.
3. The cryopreservation composition of claim 1 , wherein the gelatin has a molecular weight ranging from about 50 kDa to about 200 kDa.
4. The cryopreservation composition of claim 1 , substantially free of DMSO.
5. The cryopreservation composition of claim 1 , further comprising an amino acid, a cytokine, a lipid, a growth factor, an antibiotic, an antimycotic, a steroid hormone, a protein hormone, or a combination thereof.
6. The cryopreservation composition of claim 1 , further comprising one or more cells.
7. The cryopreservation composition of claim 6 , in a cryopreserved state.
8. The cryopreservation composition of claim 6 , wherein the cells are present at a concentration ranging from about 10 5 cells/ml to about 10 7 cells/ml.
9. The cryopreservation composition of claim 1 , comprising:
about 1% (w/v) to about 5% (w/v) of gelatin.
10. The cryopreservation composition of claim 9 , further comprising one or more cells.
11. The cryopreservation composition of claim 10 , in a cryopreserved state.
12. The cryopreservation composition of claim 9 , wherein the cells are present at a concentration ranging from about 10 5 cells/ml to about 10 7 cells/ml.
13. The cryopreservation composition of claim 1 , wherein the saccharide comprises sucrose, sorbitol, glucose, fructose, galactose, trehalose, mannose, maltose, or combinations thereof.
14. The cryopreservation composition of claim 9 , wherein the saccharide comprises sucrose, sorbitol, glucose, fructose, galactose, trehalose, mannose, maltose, or combinations thereof.
15. A kit comprising the cryopreservation composition of claim 1 .
16. A method for cryopreserving one or more cells, the method comprising the steps of:
(a) mixing the one or more cells with the cryopreservation composition of claim 1 to form a mixture, and
(b) freezing the mixture.
17. The method of claim 16 , wherein the cells are mammalian cells.
18. The method of claim 16 , wherein the cells are human, porcine, canine, equine or bovine cells.
19. The method of claim 16 , wherein the cells comprise tumor cells.
20. The method of claim 16 , wherein the cells comprise fibroblasts.
21. The method of claim 16 , wherein the cells comprise stem cells.
22. The method of claim 16 , wherein the mixture is frozen at a temperature ranging from about −70° C. to about −200° C.
23. The method of claim 16 , wherein the cells are present in the mixture at a concentration ranging from about 10 5 cells/ml to about 10 7 cells/ml.
24. The method of claim 16 , wherein the cryopreservation composition is substantially free of DMSO.
25. The method of claim 16 , further comprising the step (c) thawing the frozen mixture.
26. The method of claim 25 , wherein the cells have a post-thaw viability of at least 70%.
27. The method of claim 25 , wherein the cells have a post-thaw viability of at least 80%.