Cryopreservation of encapsulated pancreatic endoderm cells

1. A cryopreservation container comprising: a cryopreserved combination product and a cryopreservation solution, wherein the cryopreserved-combination product comprises a human pancreatic and duodenal homeobox 1 (PDX1)-positive pancreatic endoderm cell population loaded into an implantable semi-permeable encapsulation device, wherein the human PDX1-positive pancreatic endoderm cell population comprises endocrine, non-endocrine, and residual cells, and the endocrine cells are chromogranin A (CHGA)+/homeobox protein Nkx-6.1 (NKX6.1)-/PDX1- and the non-endocrine cells are CHGA−/NKX6.1+/PDX1+, and wherein the cryopreservation container comprises at least one loading port for the cryopreservation solution, and wherein the cryopreservation solution comprises about 5% to about 15% dimethyl sulfoxide (DMSO), and wherein the human PDX1-positive pancreatic endoderm cell population upon thawing and transplantation into a mammal matures in vivo into an endocrine precursor population, and the expression of neurogenin-3 (NGN3) in the PDX1-positive pancreatic endoderm cell population is lower than the expression of NGN3 in the endocrine precursor population.

2. The cryopreservation container of claim 1 , wherein the cryopreservation container is a vial, cryotube, or bag.

3. The cryopreservation container of claim 1 , wherein the container comprises less than 100 mL, less than 50 mL, less than 25 mL, less than 10 mL, less than 5 mL, or less than 3 mL of the cryopreservation solution.

4. The cryopreservation container of claim 1 , wherein the human PDX1-positive pancreatic endoderm cell population is contacted with cryopreservation solution prior to loading into the implantable semi-permeable encapsulation device.

5. The cryopreservation container of claim 1 , wherein the cryopreservation solution is equilibrated inside the implantable semi-permeable encapsulation device to about 5% to about 15% DMSO.

6. The cryopreservation container of claim 1 , wherein the combination product is pretreated prior to cryopreservation.

7. The cryopreservation container of claim 1 , wherein the human PDX1-positive pancreatic endoderm cell population is mixed with a compound, a solution, or a protein prior to cryopreservation.

8. The cryopreservation container of claim 1 , wherein the human PDX1-positive pancreatic endoderm cell population comprises human PDX1-positive pancreatic endoderm cell aggregates.

9. The cryopreservation container of claim 1 , wherein greater than about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or greater than about 95% of the human PDX1-positive pancreatic endoderm cell population can survive thawing.

10. The cryopreservation container of claim 1 , wherein the implantable semi-permeable encapsulation device comprises at least one loading port.

11. The cryopreservation container of claim 1 , wherein the cryopreserved combination product is at a temperature of about negative 90 to about negative 260 degrees Celsius.

12. The cryopreservation container of claim 1 , wherein the human PDX1-positive pancreatic endoderm cell population encapsulated in the device was contacted with a cryopreservative for at least 20 minutes prior to cryopreservation, or about 30 minutes prior to cryopreservation, or about 60 minutes prior to cryopreservation.

13. The cryopreservation container of claim 1 , wherein there is a decrease in the residual cells following thawing of the human PDX1-positive pancreatic endoderm cell population.

14. The cryopreservation container of claim 1 , wherein there is an increase in the non-endocrine cells as compared to endocrine cells following thawing of the human PDX1-positive pancreatic endoderm cell population.

15. The cryopreservation container of claim 1 , wherein there is a decrease in the residual cells compared to the non-endocrine cells following thawing of the human PDX1-positive pancreatic endoderm cell population.

16. The cryopreservation container of claim 1 , wherein there is an increase in the non-endocrine cells as compared to endocrine cells following thawing of the human PDX1-positive pancreatic endoderm cell population.

17. The cryopreservation container of claim 16 , wherein greater than 40% of the human PDX1-positive pancreatic endoderm cell population that survive thawing are endocrine cells and greater than 50% of the human PDX1-positive pancreatic endoderm cell population that survive thawing are non-endocrine cells.

18. The cryopreservation container of claim 1 , wherein the at least one loading port is sealed.