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US 12,757,893 B2Granted
Methods for cryogenic storage
AssigneeJuno Therapeutics, Inc.
InventorsSara Elizabeth Church, Jon Charles Gunther, Kathryn Pollock
Life of this patent
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Abstract
The present disclosure relates to methods and compositions and articles of manufacture, related to cryogenic storage, such as for cryogenically freezing and storing cells from a donor's blood, and methods of processing an apheresis sample.
Claims35
A method comprising cryogenically storing cells from a biological sample derived from a donor, the method comprising,
- (a) suspending a population of cells that are engineered to express a recombinant receptor in a cryopreservation medium to form a suspended composition, said cryopreservation medium comprising 0.1% to 50% dimethyl sulfoxide (DMSO) (v/v) and from 0.1% to 20% human serum albumin (HSA) (w/v), wherein the suspended composition has a cell density of about 10×106 cells/mL to about 150×106 cells/mL, and wherein the population of cells comprises at least 80% T cells and is for treating a disease or condition in the donor, and wherein the suspended composition is in a container at a surface area to volume (SA:
- V) ratio of 3 cm−1 to 6 cm−1,
- (b) cryogenically freezing the suspended composition in a controlled rate freezer comprising a chamber using a stepwise freezing profile to form cryogenically frozen cells, the stepwise freezing profile comprising a cooling step that cools the suspended composition or chamber at a rate of 1.2° C. per minute to a temperature of about −6° C., followed by a cooling step that cools the suspended composition or chamber at a rate of 25° C. per minute from about −6° C. until the suspended composition or chamber reaches a temperature of about −65° C., and
- (c) storing the cryogenically frozen cells.
1 more element
A method of cryogenically storing engineered cells, the method comprising,
- (a) suspending a population of cells that are engineered to express a recombinant receptor in a cryopreservation medium to form a suspended composition, said cryopreservation medium comprising 3% to 9% dimethyl sulfoxide (DMSO) (v/v) and from 1% to 5% human serum albumin (HSA) (w/v), wherein the suspended composition has a cell density of about 10×106 cells/mL to about 150×106 cells/mL, and wherein the population of cells comprises at least 80% T cells, and wherein the suspended composition is in a container at a surface area to volume (SA:
- V) ratio of 3 cm−1 to 6 cm−1,
- (b) cryogenically freezing the suspended composition in a controlled rate freezer comprising a chamber using a stepwise freezing profile to form cryogenically frozen cells, the stepwise freezing profile comprising a cooling step that cools the suspended composition or chamber at a rate of 1.2° C. per minute to a temperature of about −6° C., followed by a cooling step that cools the suspended composition or chamber at a rate of 25° C. per minute from about −6° C. until the suspended composition or chamber reaches a temperature of about −65° C., and
- (c) storing the cryogenically frozen cells at a temperature below −80° C. for a period of time greater than or equal to 24 hours.
1 more element
Dependencies are inferred from the claim text.