Cell freezing medium for clinical use
Provided in the present invention is a cell freezing medium for clinical use. In particular, the cell freezing medium of the present invention comprises the following components: (1) human albumin; (2) cryoprotectant: the cryoprotectant comprises a combination of one or more of dimethyl sulfoxide, glycerol, and ethylene glycol; (3) a saline buffer; wherein the salt buffer is a solution containing Na + , K + , Mg + , Cl − , and CH 3 COO − ions; (4) a vitamin; and (5) an amino acid, wherein the human albumin concentration is 1%-20% (w/v). The cell, after long-term cryopreservation with the freezing medium of the present invention, has a high viability, and the cellular efficiency maintains a high uniformity. The grade of purity of the freezing medium of the present invention is the pharmaceutical grade or USP grade; and the freezing medium is safe and reliable for clinical use, and can be used or conventional adherent and suspension cells.
1. A cell cryopreservation solution for long-term storage of a mammalian stem cell, comprising:
(a) an albumin; and
(b) amino acids; wherein the amino acids consist of a combination of glycine and arginine or a combination of leucine and phenylalanine.
2. The cell cryopreservation solution of claim 1 , further comprising a cryoprotectant comprising one or more of dimethyl sulfoxide, glycerin, and ethylene glycol.
3. The cell cryopreservation solution of claim 1 , further comprising a salt buffer comprising one or more of Na + , K + , Mg 2+ , Cl − and CH3COO − ions.
4. The cell cryopreservation solution of claim 1 , further comprising a vitamin.
5. The cell cryopreservation solution of claim 1 , wherein the albumin is a human albumin.
6. The cell cryopreservation solution of claim 1 , wherein the albumin is present at a concentration of 1%-20% weight by volume (w/v).
7. The cell cryopreservation solution of claim 5 , wherein the human albumin is selected from the group consisting of a plasma extracted human albumin, a recombinant human albumin, and a combination thereof.
8. The cell cryopreservation solution of claim 1 , wherein the amino acids are independently present at a concentration of 0.1 mol/L-0.5 mol/L.
9. The cell cryopreservation solution of claim 2 , wherein the cryoprotectant is present at a concentration of 0.4 mol/L-2.2 mol/L.
10. The cell cryopreservation solution of claim 3 , wherein concentrations of Na + , K + , Mg 2+ , Cl − and CH3COO − in the salt buffer are one or more concentrations selected from the group consisting of:
a concentration of the Na + ions being 0.1-0.2 mol/L;
a concentration of the K + ions being 5-10 mmol/L;
a concentration of the Mg 2+ ions being 1.5-5 mmol/L;
a concentration of the Cl − ions being 0.1-0.3 mol/L; and
a concentration of the CH3COO − ions being 40-60 mmol/L.
11. The cell cryopreservation solution of claim 4 , wherein the vitamin is present at a concentration of 50 mmol/L-150 mmol/L.
12. The cell cryopreservation solution of claim 4 , wherein the vitamin is selected from the group consisting of vitamin A, vitamin B, vitamin C, vitamin E, vitamin K, and combinations thereof.
13. The cell cryopreservation solution of claim 12 , wherein the vitamin is vitamin C or vitamin E.
14. A mammalian cell mixture, comprising a mammalian cell and the cell cryopreservation solution of claim 1 .
15. The mammalian cell mixture of claim 14 , wherein the mammalian cell is a mesenchymal stem cell or a CAR-T cell.
16. A method for cryopreservation of a mammalian cell, comprising:
(i) mixing the mammalian cell to be cryopreserved with the cell cryopreservation solution of claim 1 , thereby obtaining a mammalian cell mixture;
(ii) cooling the mammalian cell mixture obtained in step (i) and storing the mammalian cell mixture.
17. A kit for cell cryopreservation, comprising the cell cryopreservation solution of claim 1 .