Cryopreservative compositions and methods
This disclosure describes a cryopreservative composition and methods for storing cells. Generally, the cryopreservative composition includes a sugar component and a sugar alcohol component, and is effective for storing and recovering cells without requiring dimethyl sulfoxide (DMSO).
1. A method of cryopreserving a cell, the method comprising:
adding a cell to a cryopreservative composition comprising:
a sugar component; and
a sugar alcohol component;
wherein total concentration of sugar alcohol components in the composition is between 0.1 M and 2 M, with the proviso that the composition includes less than a cryopreservative amount of dimethyl sulfoxide (DMSO); and
freezing the composition.
2. The method of claim 1 , further comprising:
storing the frozen composition at a temperature below 0° C.; and
thawing the composition,
wherein the cell remains viable after thawing.
3. The method of claim 1 , wherein the sugar component is provided at a concentration of 0.1 mM to 300 mM.
4. The method of claim 1 , wherein the sugar component is provided at a concentration of 10 mM to 300 mM.
5. The method of claim 1 , wherein the sugar component comprises trehalose, sucrose, lactose, maltose, or a combination thereof.
6. The method of claim 1 , wherein the sugar alcohol component is provided at a concentration of 0.1 M to 1.4 M.
7. The method of claim 1 , wherein the sugar alcohol component is provided at a concentration of 0.1 M to 0.6 M.
8. The method of claim 1 , wherein the sugar alcohol component comprises sorbitol, ethylene glycol, inositol, xylitol, mannitol, glycerol, arabitol, erythritol, ribitol, or a combination thereof.
9. The method of claim 1 , wherein the sugar alcohol component comprises glycerol at a concentration of 0.6 M to 1.4 M.
10. The method of claim 1 , further comprising an additive component comprising an amino acid.
11. The method of claim 1 , wherein less than a cryopreservative amount of dimethyl sulfoxide (DMSO) is an amount of DMSO that provides no more cryoprotection than the remaining components of the composition without the DMSO.
12. The method of claim 1 , wherein freezing the composition comprises at least one round of cooling, re-warming, and further cooling.
13. A method of cryopreserving a cell, the method comprising:
adding a cell to a cryopreservative composition comprising:
a sugar component comprising a disaccharide; and
a sugar alcohol component;
wherein total concentration of sugar alcohol components in the composition is no more than 2 M, with the proviso that the composition includes less than a cryopreservative amount of dimethyl sulfoxide (DMSO); and
freezing the composition.
14. The method of claim 13 , further comprising:
storing the frozen composition at a temperature below 0° C.;
thawing the composition;
removing the cell from the thawed composition; and
culturing the cell under conditions effective for the cell to remain viable.
15. The method of claim 13 , wherein the sugar component is provided at a concentration of 0.1 mM to 300 mM.
16. The method of claim 13 , wherein the sugar component comprises trehalose, sucrose, lactose, maltose, or a combination thereof.
17. The method of claim 13 , wherein the sugar alcohol component comprises glycerol, sorbitol, ethylene glycol, inositol, xylitol, mannitol, arabitol, erythritol, ribitol, or a combination thereof.
18. A method of cryopreserving a cell, the method comprising:
adding a cell to a cryopreservative composition comprising:
a sugar component comprising a disaccharide, the sugar component being provided at a concentration of up to 300 mM;
a sugar alcohol component at a concentration of 0.1 M to 2 M; and
less than a cryopreservative amount of dimethyl sulfoxide (DMSO); and
freezing the composition.
19. The method of claim 18 , further comprising:
storing the frozen composition at a temperature below 0° C.;
thawing the composition;
removing the cell from the thawed composition; and
culturing the cell under conditions effective for the cell to remain viable.
20. The method of claim 18 , wherein the sugar component comprises trehalose, sucrose, lactose, maltose, or a combination thereof, and
wherein the sugar alcohol component comprises glycerol, sorbitol, ethylene glycol, inositol, xylitol, mannitol, arabitol, erythritol, ribitol, or a combination thereof.