Compositions and methods of cryopreserving cells

Patent metadata

Field	Value
Publication number	US-12433289-B2
Application number	US-202117153458-A
Country	US
Kind code	B2
Filing date	Jan 20, 2021
Priority date	Jan 21, 2020
Publication date	Oct 7, 2025
Grant date	Oct 7, 2025

Abstract

The present disclosure provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: dimethyl sulfoxide (DMSO), disaccharide, human serum, and IL-7 and/or IL-15. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w/v % and 10 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and between about 10 w/v % and 90 w/v % human serum. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w/v % and 10 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and between about 0.5 w/v % and 30 w/v % human serum albumin.

First claim

We claim:

1. A composition comprising:
mammalian immune cells that are primary cells or derived from induced pluripotent stem cells; and
a cryopreservation medium comprising: about 5 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and human serum albumin (HSA), wherein the disaccharide is sucrose.

2. The composition of claim 1 , further comprising one or more inorganic salts.

3. The composition of claim 1 , wherein the human serum albumin (HSA) is present at a final concentration of about 2.5% (w/v).

4. The composition of claim 1 , wherein the mammalian immune cells are lymphocytes.

5. The composition of claim 4 , wherein the mammalian immune cells are T cells.

6. The composition of claim 4 , wherein the mammalian immune cells are natural killer (NK) cells.

7. The composition of claim 4 , wherein the mammalian immune cells are induced pluripotent cell (iPSC)-derived lymphocytes.

8. The composition of claim 7 , wherein the induced pluripotent cell (iPSC)-derived lymphocytes are T cells.

9. The composition of claim 7 , wherein the induced pluripotent cell (iPSC)-derived lymphocytes are natural killer (NK) cells.

10. The composition of claim 1 , comprising the mammalian immune cells at a concentration of about 1×10 6 to about 3×10 8 cells/mL.

11. The composition of claim 1 , wherein the mammalian immune cells are activated without cytokines.

12. The composition of claim 1 , wherein the mammalian immune cells are primary cells directly isolated from a non-patient donor.

13. The composition of claim 1 , wherein the mammalian immune cells are primary cells directly isolated from a patient.

14. The composition of claim 1 , wherein the mammalian immune cells are genetically modified.

15. The composition of claim 1 , wherein the mammalian immune cells are iCART cells.

16. The composition of claim 1 , comprising between 0.5 w/v % and 30 w/v % human serum albumin (HSA).

17. A composition comprising:
mammalian immune cells that are primary cells or derived from induced pluripotent stem cells; and
a cryopreservation medium comprising: about 5 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and human serum albumin (HSA), further comprising cytokines, wherein the cytokines are IL-7 and IL-15.

18. The composition of claim 17 , wherein IL-7 is present at a final concentration of between about 1 ng/mL and 50 ng/mL.

19. The composition of claim 17 , wherein IL-15 is present at a final concentration of between about 1 ng/ml and 50 ng/mL.

20. A composition comprising:
mammalian immune cells that are primary cells or derived from induced pluripotent stem cells; and
a cryopreservation medium comprising: about 5 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and human serum albumin (HSA), further comprising a mammalian cell culture medium.

21. The composition of claim 20 , wherein the mammalian cell culture medium is present at between about 10 w/v % and 90 w/v %.

22. A composition comprising:
mammalian immune cells that are primary cells or derived from induced pluripotent stem cells; and
a cryopreservation medium comprising: about 5 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and human serum albumin (HSA), further comprising one or more amino acids.

Assignees

• Takeda Pharmaceuticals Co

Inventors

• Cao Lan
• Chang Hui-Hsin
• Hu Jianxin
• Shi Xi
• Nishimoto Yutaka

Classifications

• A01N1/126

  Physiologically active agents, e.g. antioxidants or nutrients · CPC title

• A01N1/162

  Temperature processes, e.g. following predefined temperature changes over time · CPC title

• A01N1/125 Primary

  Freeze protecting agents, e.g. cryoprotectants or osmolarity regulators · CPC title

• A01N1/10 Primary

  Preservation of living parts · CPC title