title: Long-term in vitro human pancreatic islet culture using three-dimensional microfabricated scaffolds.
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.
authors.affiliation: Department of Biomedical Engineering, Faculty of Medicine, McGill University, 3775 University Street, Montreal (QC), Canada. jamal.daoud@mail.mcgill.ca
authors.forename: Jamal T
authors.initials: JT
authors.lastname: Daoud
authors.forename: Maria S
authors.initials: MS
authors.lastname: Petropavlovskaia
authors.forename: Jason M
authors.initials: JM
authors.lastname: Patapas
authors.forename: Christian E
authors.initials: CE
authors.lastname: Degrandpré
authors.forename: Robert W
authors.initials: RW
authors.lastname: Diraddo
authors.forename: Lawrence
authors.initials: L
authors.lastname: Rosenberg
authors.forename: Maryam
authors.initials: M
authors.lastname: Tabrizian
mesh_terms: D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D054457:Tissue Scaffolds