Directly injectable formulations which provide enhanced cryoprotection of cell products
View Patent ↗This invention provides compositions and methods for cryoprotection of recombinant live cancer cells. Specifically, an improved cryoprotective medium is provided which includes a hydroxyethyl starch and/or derivative thereof alone or in combination with either DMSO or glycerol.
1. A suspension of recombinant live cancer cells comprising a plurality of recombinant live cancer cells in a cryoprotective medium, wherein the cryoprotective medium comprises about 5% by weight to about 22% by weight of a hydroxyethyl starch (HES) and about 5% by weight to about 10% by weight of DMSO, and
wherein the recombinant live cancer cells express granulocyte-macrophage colony stimulating factor (GM-CSF).
2. The suspension of claim 1 , further comprising from about 0% by weight to about 10% by weight human serum albumin (HSA).
3. The suspension according to claim 2 , wherein the cryoprotective medium comprises about 8% by weight HES, about 2% by weight HSA, and about 5% by weight DMSO.
4. A cryopreserved, frozen suspension of recombinant live cancer cells, comprising a plurality of recombinant live cancer cells in a cryoprotective medium, wherein the cryoprotective medium comprises about 5% by weight to about 22% by weight of a hydroxyethyl starch (HES) and about 5% by weight to about 10% by weight of DMSO, and
wherein the recombinant live cancer cells express granulocyte-macrophage colony stimulating factor (GM-CSF).
5. The suspension according to claim 4 , further comprising from about 0% by weight to about 10% by weight human serum albumin (HSA).
6. The suspension according to claim 5 , wherein the cryoprotective medium comprises about 8% by weight HES, about 2% by weight HSA, and about 5% by weight DMSO.
7. The suspension according to any one of claims 1 - 3 or 4 - 6 , wherein the HES comprises HES: (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is from about 0.60-0.80; (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from about 0.40-0.60; or a combination of (a) and (b).
8. The suspension according to claim 7 , wherein the HES comprises HES: (a) etherified with hydroxyethyl groups wherein the degree of molecular substitution is about 0.70; (b) etherified with hydroxyethyl groups, wherein the degree of molecular substitution is from 0.4 to about 0.5; or a combination of (a) and (b).
9. The suspension of according to any one of claims 1 - 3 or 4 - 6 , wherein the HES comprises HES with: (a) a molecular weight of from about 420 to about 480 kDa; (b) a molecular weight of from about 200 to about 290 kDa; or a combination of (a) and (b).
10. The suspension according to any one of claims 1 - 3 and 4 - 6 , wherein the plurality of recombinant live cancer cells comprise an introduced heterologous nucleic acid coding sequence for GM-CSF.
11. The suspension according to claim 10 , wherein the recombinant live cancer cells express an additional cytokine or a costimulatory molecule.
12. The suspension according to any one of claims 1 - 3 or 4 - 6 , wherein the recombinant live cancer cell is selected from the group consisting of a lung cancer cell, a pancreatic cancer cell, a prostate cancer cell, a kidney cancer cell, a myeloma cell, and a leukemic cell.
13. The suspension according to any one of claims 1 - 3 or 4 - 6 , wherein the recombinant live cancer cell suspension comprises patient-derived tumor cells.
14. The suspension according to claim 13 , wherein said recombinant live cancer cell is autologous, allogeneic or bystander cells.