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. a human albumin;
b. a cryoprotectant comprising one or more of dimethyl sulfoxide, glycerin, and ethylene glycol;
c. a salt buffer comprising Na + , K + , Mg 2+ , Cl − , and CH 3 COO − ions;
d. a vitamin; and
e. amino acids;
wherein the human albumin is present at a concentration of 1%-20% weight by volume (w/v),
the amino acids consist of a combination of glycine and arginine or a combination of leucine
and phenylalanine, and
the amino acids are independently present at a concentration of 0.1 mol/L-0.5 mol/L.
2. The cell cryopreservation solution of claim 1 , wherein the cryoprotectant is present at a concentration of 0.4 mol/L-2.2 mol/L.
3. The cell cryopreservation solution of claim 1 , 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 CH 3 COO − ions being 40-60 mmol/L.
4. The cell cryopreservation solution of claim 1 , wherein the vitamin is present at a concentration of 50 mmol/L-150 mmol/L.
5. The cell cryopreservation solution of claim 1 , wherein the human albumin is selected from the group consisting of a plasma extracted human albumin, a recombinant human albumin, and a combination thereof.
6. The cell cryopreservation solution of claim 1 , wherein the vitamin is selected from the group consisting of vitamin A, vitamin B, vitamin C, vitamin E, vitamin K, and combinations thereof.
7. The cell cryopreservation solution of claim 1 , wherein the vitamin is vitamin C or vitamin E.
8. A mammalian cell mixture, comprising:
a mammalian cell and the cell cryopreservation solution of claim 1 .
9. 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 in liquid nitrogen.
10. A kit for cell cryopreservation, comprising the cell cryopreservation solution of claim 1 .
11. The mammalian cell mixture of claim 8 , wherein the mammalian cell is a mesenchymal stem cell or a CAR-T cell.