Long-term in vitro human pancreatic islet culture using three-dimensional microfabricated scaffolds

Biomaterials. 2011 Feb;32(6):1536-42. doi: 10.1016/j.biomaterials.2010.10.036. Epub 2010 Nov 19.

Abstract

Human pancreatic islet in vitro culture is very challenging and requires the presence of various extra cellular matrix (ECM) components in a three-dimensional environment, which provides mechanical and biological support. The development of such an environment is vital in providing favourable conditions to preserve human islets in long-term culture. In this study, we investigated the effects of human islet culture within various three-dimensional environments; collagen I gel, collagen I gel supplemented with ECM components fibronectin and collagen IV, and microfabricated scaffold with ECM-supplemented gel. The cultured human islets were analyzed for functionality, gene expression and hormone content following long-term in vitro culture. It was clear the incorporation of ECM components within the three-dimensional support improved prolonged culture. However, long-term and highly uniform human islet culture within a microfabricated scaffold, with controlled pore structures, coupled with the presence of ECM components, displayed an insulin release profile similar to freshly isolated islets, yielding a stimulation index of approximately 1.8. Moreover, gene expression was markedly increased for all pancreatic genes, giving a approximately 50-fold elevation of insulin gene expression with respect to suspension culture. The distribution and presence of pancreatic hormones was also highly elevated. These findings provide a platform for the long-term maintenance and preservation of human pancreatic islets in vitro.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Extracellular Matrix / chemistry
  • Fluorescent Antibody Technique
  • Humans
  • Islets of Langerhans / cytology*
  • Islets of Langerhans / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tissue Scaffolds*