Cryopreserved endocrine cells that express chromogranin a

Abstract

A human immature endocrine cell population and methods for making an immature endocrine cell population are provided. Specifically, immature beta cells and methods for production of immature beta cells are described. Immature beta cells co-express INS and NKX6.1 and are uni-potent and thereby develop into mature beta cells when implanted in vivo. The mature beta cells in vivo are capable of producing insulin in response to glucose stimulation.

1 . A method of producing mature beta cells in a subject, comprising transplanting into the subject an in vitro cell culture comprising human cells, wherein at least 10% of the human cells are unipotent immature beta cells that co-express INS and NKX6.1 and which do not substantially express NGN3, thereby producing mature beta cells in the subject. 
     
     
         2 . The method of  claim 1 , wherein at least 30% of the human cells are unipotent immature beta cells that co-express INS and NKX6.1 and which do not substantially express NGN3. 
     
     
         3 . The method of  claim 1 , wherein the cell culture is in a semipermeable encapsulation device. 
     
     
         4 . The method of  claim 1 , wherein the cell culture is in a perforated encapsulation device. 
     
     
         5 . The method of  claim 1 , wherein the unipotent immature beta cells are in cell aggregates. 
     
     
         6 . The method of  claim 5 , wherein the cell aggregates are 50 to 600 microns in size. 
     
     
         7 . The method of  claim 5 , wherein the cell aggregates are 50 to 250 microns in size. 
     
     
         8 . The method of  claim 1 , wherein the unipotent immature beta cells further express pancreatic duodenal homeobox factor-1 (PDX-1). 
     
     
         9 . The method of  claim 1 , wherein the unipotent immature beta cells further express MAFB. 
     
     
         10 . The method of  claim 1 , wherein the unipotent immature beta cells are capable of maturing into mature beta cells. 
     
     
         11 . The method of  claim 1 , wherein the cell culture further comprises Chromogranin A (CHGA)-negative cells. 
     
     
         12 . The method of  claim 1 , wherein the subject has diabetes. 
     
     
         13 . A method of producing insulin in a subject, comprising transplanting into the subject an in vitro cell culture comprising human cells, wherein at least 10% of the human cells are unipotent immature beta cells that co-express INS and NKX6.1 and which do not substantially express NGN3, thereby producing insulin in the subject. 
     
     
         14 . The method of  claim 13 , wherein at least 30% of the human cells are unipotent immature beta cells that co-express INS and NKX6.1 and which do not substantially express NGN3. 
     
     
         15 . The method of  claim 13 , wherein the cell culture is in a semipermeable encapsulation device. 
     
     
         16 . The method of  claim 13 , wherein the cell culture is in a perforated encapsulation device. 
     
     
         17 . The method of  claim 13 , wherein the unipotent immature beta cells are in cell aggregates. 
     
     
         18 . The method of  claim 17 , wherein a) the cell aggregates are 50 to 600 microns in size or b) the cell aggregates are 50 to 250 microns in size. 
     
     
         19 . The method of  claim 13 , wherein a) the unipotent immature beta cells further express pancreatic duodenal homeobox factor-1 (PDX-1) and/or b) the unipotent immature beta cells further express MAFB. 
     
     
         20 . The method of  claim 13 , wherein the subject has diabetes.