Cryopreservation, hibernation and room temperature storage of encapulated pancreatic endoderm cell aggregates

Abstract

Disclosed herein are methods for cryopreserving, hibernation and room temperature storage of PEC aggregates, implantable semipermeable devices and the VC combination product.

1 . A method for cryopreserving an encapsulated cell population, said method comprising:
 a. obtaining a cell population to be cryopreserved;   b. loading the cell population into an implantable semi-permeable encapsulation device thereby making an encapsulated cell population;   c. contacting the encapsulated cell population with a cryopreservative thereby cryopreserving the encapsulated cell population.   
     
     
         2 . A method for producing insulin in vivo in a mammal, said method comprising:
 a. obtaining an in vitro human pancreatic cell aggregate population;   b. loading the pancreatic cell aggregate population into an implantable semi-permeable encapsulation device thereby making an encapsulated pancreatic cell population;   c. contacting the pancreatic cell population with cryopreservative thereby cryopreserving the encapsulated pancreatic cell population;   d. thawing the encapsulated pancreatic cell population;   e. implanting the encapsulated pancreatic cell population into a mammalian host; and   f. maturing the encapsulated pancreatic cell population in vivo to form a mature cell population comprising of endocrine and acinar cells, wherein at least some of the endocrine cells are insulin secreting cells that produce insulin in response to glucose stimulation in vivo, thereby producing insulin in vivo to the mammal.   
     
     
         3 . The method of  claim 1 , wherein the cell population to be cryopreserved is PDX1 positive pancreatic endoderm cells. 
     
     
         4 . The method of  claim 3 , wherein the PDX1 positive pancreatic endoderm cells are pancreatic endoderm cells. 
     
     
         5 . The method of  claim 1 , wherein the cell population to be cryopreserved is contacted with a cryopreservative prior to loading into the device. 
     
     
         6 . The method of  claim 1 , wherein the encapsulated cell population is shipped to the implantation site in a cryopreserved state. 
     
     
         7 . The method of  claim 1 , wherein the encapsulated cell population is at a temperature range of negative 90 to negative 260 degrees Celsius. 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 1 , further comprising thawing the encapsulated cell population, wherein the survival rate for the thawed encapsulated cell population is 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%. 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . The method of  claim 1 , wherein the device comprises at least one loading port. 
     
     
         15 . (canceled) 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . A cryopreserved PDX1 positive pancreatic endoderm population. 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . The method of  claim 1 , wherein the encapsulated cell population is pancreatic endoderm cell (PEC) aggregates. 
     
     
         29 . The cryopreserved PDX1 positive pancreatic endoderm population of  claim 19  wherein the PDX1 positive pancreatic endoderm population is in contact with cryopreservative. 
     
     
         30 . The cryopreserved PDX1 positive pancreatic endoderm population of  claim 19  wherein the PDX1 positive pancreatic endoderm population is PEC aggregates. 
     
     
         31 . The method of  claim 2 , 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 encapsulated pancreatic cell population survive thawing. 
     
     
         32 . The cryopreserved PDX1 positive pancreatic endoderm population of  claim 19  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 PDX1 positive pancreatic endoderm population survive thawing. 
     
     
         33 . The method of  claim 2 , wherein the device comprises at least one loading port. 
     
     
         34 . The cryopreserved PDX1 positive pancreatic endoderm population of  claim 19  wherein the PDX1 positive pancreatic endoderm population is loaded into an implantable semi-permeable encapsulation device. 
     
     
         35 . The cryopreserved PDX1 positive pancreatic endoderm population of  claim 34  wherein the device comprises at least one loading port. 
     
     
         36 . The method of  claim 2 , wherein the encapsulated pancreatic cell population is at a temperature range of negative 90 to negative 260 degrees Celsius.