Method for the cryopreservation of cells for therapeutic purposes
View Patent ↗The present invention relates to a composition comprising, in a physiologically acceptable medium: a) at least one saccharide, b) at least one amino acid, c) DMSO or at least one C3-C5 alkanediol, d) at least one antioxidant, and e) cells for therapeutic purposes, said composition having a pH between 7.0 and 8.5, preferably between 7.0 and 8.3. It also relates to a method for the cryopreservation of at least one sample of cells for therapeutic purposes, comprising the following steps: i) mixing the sample of cells for therapeutic purposes with ingredients a) to d) above and a physiologically acceptable medium, so as to obtain a composition having a pH between 7.0 and 8.5, preferably between 7.0 and 8.3, then ii) freezing the composition obtained in step i).
1. A composition comprising, in a physiologically acceptable medium:
a) at least one saccharide,
b) at least one amino acid,
c) DMSO or at least one C3-C5 alkanediol,
d) at least one antioxidant,
e) platelet lysate, and
f) cells for therapeutic purposes,
said composition having a pH between 7.0 and 8.5,
wherein the cells for therapeutic purposes are natural or genetically modified T cells or myoblasts.
2. The composition according to claim 1 , wherein its pH is between 7.0 and 8.3.
3. The composition according to claim 1 , wherein the saccharide is a monosaccharide, disaccharide, or trisaccharide.
4. The composition according to claim 1 , wherein the amino acid is selected from glutamine, alanyl-glutamine, tryptophan, lysine, methionine, phenylalanine, threonine, valine, leucine and isoleucine, arginine, histidine, tyrosine, cysteine, and mixtures thereof.
5. The composition according to claim 1 , wherein the amino acid is cysteine.
6. The composition according to claim 1 , wherein the cysteine is in the form of cystine, said cystine being a cysteine dimer.
7. The composition according to claim 1 , further comprising at least one vitamin.
8. The composition according to claim 1 , wherein the vitamin is selected from vitamins B1, B2, B4, B5, B6, B7, B9, PP, and mixtures thereof.
9. The composition according to claim 1 , wherein the C3-C5 alkanediol is selected from the linear alkanes comprising from 3 to 5 carbon atoms and 2 hydroxyl groups.
10. The composition according to claim 1 , wherein the C3-C5 alkanediol is propylene glycol.
11. The composition according to claim 1 , wherein the antioxidant is glutathione, vitamin C, vitamin E, vitamin A, L-cysteine, or coenzyme Q10.
12. The composition according to claim 1 , wherein the antioxidant is glutathione.
13. The composition according to claim 1 , further comprising human albumin.
14. The composition according to claim 1 , further comprising at least one bicarbonate salt.
15. The composition according to claim 1 , wherein myoblasts are human ones.
16. The composition according to claim 1 , wherein the physiologically acceptable medium is an aqueous medium comprising electrolytes, and wherein the composition comprises:
a) glucose,
b) a mixture of glutamine, alanyl-glutamine, tryptophan, lysine, methionine, phenylalanine, threonine, valine, leucine, isoleucine, arginine, histidine, tyrosine, and cysteine,
c) DMSO or at least one C3-C5 alkanediol,
d) glutathione, and
e) cells for therapeutic purposes.
17. A method for the cryopreservation of at least one sample of cells for therapeutic purposes, comprising:
i) mixing the sample of cells for therapeutic purposes with:
a) at least one saccharide,
b) at least one amino acid,
c) DMSO or at least one C3-C5 alkanediol,
d) at least one antioxidant,
e) platelet lysate, and
a physiologically acceptable medium, to obtain a composition having a pH between 7.0 and 8.5, wherein said cells are natural or genetically modified T cells or myoblasts, and
ii) freezing the composition obtained in step i).
18. The method according to claim 17 , wherein the pH is between 7.0 and 8.3.
19. The method according to claim 17 , wherein the freezing ii) is carried out to a temperature between −100° C. and −180° C.
20. The method according to claim 17 , wherein the freezing step ii) is carried out by placing the composition obtained in step i) in a container immersed in a mixture of isopropyl alcohol at +4° C., then bringing the whole to a temperature between −70° C. and −100° C.
21. A method for the cryopreservation of at least one sample of cells for therapeutic purposes, said method comprising mixing said sample of cells with a composition comprising, in a physiologically acceptable medium:
a) at least one saccharide,
b) at least one amino acid,
c) DMSO or at least one C3-C5 alkanediol,
d) at least one antioxidant,
e) platelet lysate, and having a pH between 7.0 and 8.5,
wherein said cells are natural or genetically modified T cells or myoblasts.
22. The cryopreservation method according to claim 21 , wherein the composition has a pH between 7.0 and 8.3.