Coculture With Ischemia/Reperfusion-Preconditioned Hepatocytes Improves Islet Function and Survival

Transplant Proc. 2018 Dec;50(10):3887-3894. doi: 10.1016/j.transproceed.2018.07.007. Epub 2018 Jul 7.

Abstract

In clinical islet transplantation, hepatic ischemia and insufficient neovascularization of transplanted islets are barriers to islet survival and function. However, hepatocytes have a potency to protect themselves against ischemia. We hypothesized that ischemia/reperfusion preconditioning (IRP) of hepatocytes might beneficially affect islet cells in a coculture system. Primary islets were cocultured with primary hepatocytes, and hepatocyte IRP was conducted by subjecting cells to hypoxic conditions for single 15-minute/30-minute hypoxia, or 2 tandem 15-minute/30-minute hypoxic treatments (hypoxic-normoxic-hypoxic). We show that gene expression levels of insulin-like growth factor 1 (IGF-1), hepatocyte growth factor (HGF), transforming growth factor-α (TGF-α), and TGF-β1 in hepatocytes were increased by IRP. IRP hepatocytes secreted hepatocyte growth factor and insulin-like growth factor-1. Coculture of islets with IRP hepatocytes enhanced islet insulin secretion in glucose challenge test and expression of the survival-related gene Bcl-2 and the regenerating gene-1α (Reg-1α). Islets cocultured with the 30-minute double-IRP hepatocytes displayed significantly higher viability in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and terminal deoxynucleotidyl transferase dUTP nick end labeling stain compared with that of islets subjected to 30 minutes of hypoxia. These results suggest that islet coculture with IRP hepatocytes can improve islet survival and insulin secretion.

MeSH terms

  • Animals
  • Cell Survival
  • Coculture Techniques
  • Hepatocyte Growth Factor / metabolism
  • Hepatocytes / cytology*
  • Hepatocytes / metabolism
  • Insulin / metabolism
  • Insulin-Like Growth Factor I / metabolism
  • Ischemic Preconditioning / methods*
  • Islets of Langerhans / cytology*
  • Islets of Langerhans Transplantation / methods*

Substances

  • HGF protein, human
  • IGF1 protein, human
  • Insulin
  • Hepatocyte Growth Factor
  • Insulin-Like Growth Factor I