Functionality and antidiabetic utility of β- and L-cell containing pseudoislets

Abstract

Unavailability of tissue and poor engraftment remain significant obstacles to clinical islet transplantation. Here, the therapeutic potential of pseudoislets generated from the insulin and GLP-1 releasing cell-lines MIN6 and GLUTag was investigated. Glucose and other secretagogues evoked 1.3-5.7 fold increases in insulin secretion from both pseudoislet types. Secretion expressed in relation to basal values did not greatly differ between configurations. Exposure of both types of pseudoislets to ninhydrin, H2O2, streptozotocin or cytokine cocktails decreased viability and increased apoptosis. However, combined pseudoislets exhibited enhanced resistance (1.2-1.7 fold increased LD50, 1.2-1.4 fold decreased apoptosis). Implantation of pseudoislets into streptozotocin-diabetic SCID mice precipitated cell masses containing immunoreactive insulin and GLP-1. Implantation of both pseudoislet types was associated with significant reductions in blood glucose, increased plasma insulin, greater bodyweight, decreased polydipsia and improved glucose tolerance. These changes greatly exaggerated in MIN6 pseudoislet recipients, with mice becoming severely hypoglycaemic. In contract, combined pseudoislet recipients achieved tempered restoration of normoglycaemia and exhibited increased plasma GLP-1, decreased plasma and pancreatic glucagon, increased pancreatic insulin and enhancements in islet β:α cells and the ratio of Ki67: TUNEL positive β-cells. MIN6 pseudoislet implantation increased islet β:α cell ratio but did not affect β-cell proliferation or hormone content. Our observations highlight the potential of combining insulin and GLP-1 cell therapy using heterotypic pseudoislets.

Keywords: Cell therapy; Diabetes; GLP-1; GLUTag; Insulin; MIN6; Pseudoislets.