Cryopreservation and storage of an implantable encapsulation device loaded with an endocrine cell population

Abstract

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

We claim: 
     
         1 . A cryopreservation container, comprising: a cryopreserved combination product and a cryopreservation solution,
 wherein the cryopreserved combination product comprises a human endocrine cell population loaded into an implantable semi-permeable encapsulation device,   wherein the human endocrine cell population is chromogranin A (CHGA)+, insulin (INS)+, and homeobox protein Nkx-6.1 (NKX6.1)+,   wherein the cryopreservation container comprises at least one loading port for the cryopreservation solution,   wherein the cryopreservation solution comprises about 5% to about 15% dimethyl sulfoxide (DMSO), and   wherein the human endocrine cells, upon thawing and transplantation into a human, mature in vivo into pancreatic islet cells.   
     
     
         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 endocrine cell population is contacted with a cryopreservation solution prior to loading into the implantable semi-permeable encapsulation device. 
     
     
         5 . The cryopreservation container of  claim 1 , wherein the cryopreservation solution comprises about 10% DMSO. 
     
     
         6 . The cryopreservation container of  claim 1 , wherein the combination product is pretreated prior to cryopreservation by incubating the combination product in a medium comprising an antibiotic. 
     
     
         7 . The cryopreservation container of  claim 1 , wherein prior to cryopreservation, the human endocrine cell population is mixed with a compound, a solution, an agent, an additive, or a protein at a concentration sufficient to protect cell membranes of the human endocrine cell population from damage post-thaw. 
     
     
         8 . The cryopreservation container of  claim 1 , wherein the human endocrine cell population comprises human endocrine cell population 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 endocrine 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 endocrine cell population is encapsulated in the semi-permeable encapsulation device, and wherein the human endocrine cell population 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 8 , wherein the human endocrine cell population is encapsulated in the semi-permeable encapsulation device, and wherein the human endocrine cell population was contacted with a cryopreservative for about 30 to about 60 minutes, for at least 90 minutes, at least 120 minutes, about 2 to about 24 hours, or at least 24 hours. 
     
     
         14 . The cryopreservation container of  claim 1 , wherein the at least one loading port of the cryopreservation container is sealed. 
     
     
         15 . 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 aggregate population loaded into two or more cell chambers of an implantable semi-permeable encapsulation device and each of the two or more cell chambers are laterally connected by a cell-free zone,   wherein the human PDX1-positive pancreatic endoderm cell aggregate population comprises endocrine, non-endocrine, and residual cells, 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+,   wherein the cryopreservation container further comprises at least one loading port for the cryopreservation solution,   wherein the cryopreservation solution comprises about 5% to about 15% dimethyl sulfoxide (DMSO), and   wherein the human PDX1-positive pancreatic endoderm cell aggregate 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 aggregate population is lower than the expression of NGN3 in the endocrine precursor population.