Abstract

Clinical islet transplantation is a promising treatment for patients with type 1 diabetes. However, pancreatic islets vary in size and shape affecting their survival and function after transplantation because of mass transport limitations. To reduce diffusion restrictions and improve islet cell survival, the generation of islets with optimal dimensions by dispersion followed by reassembly of islet cells, can help limit the length of diffusion pathways. This study describes a microwell platform that supports the controlled and reproducible production of three-dimensional pancreatic cell clusters of human donor islets. We observed that primary human islet cell aggregates with a diameter of 100–150 μm consisting of about 1000 cells best resembled intact pancreatic islets as they showed low apoptotic cell death (<2%), comparable glucose-responsiveness and increasing PDX1, MAFA and INSULIN gene expression with increasing aggregate size. The re-associated human islet cells showed an a-typical core shell configuration with beta cells predominantly on the outside unlike human islets, which became more randomized after implantation similar to native human islets. After transplantation of these islet cell aggregates under the kidney capsule of immunodeficient mice, human C-peptide was detected in the serum indicating that beta cells retained their endocrine function similar to human islets. The agarose microwell platform was shown to be an easy and very reproducible method to aggregate pancreatic islet cells with high accuracy providing a reliable tool to study cell–cell interactions between insuloma and/or primary islet cells.

Highlights

  • Allogeneic islet transplantation offers a promising therapy for patients with type 1 diabetes, this method is still inefficient because of considerable islet loss shortly after transplantation [1, 2]

  • Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine

  • INS-1E cell aggregates obtained by conventional suspension culture on ultra-low attachment plates showed a large size distribution with an average aggregate diameter of 88 Æ 49 lm, whereas controlled aggregation of 500 cells per well in our microwell platform resulted in uniformly sized aggregates of 93 lm (SD Æ 16; Fig. 2E)

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Summary

Introduction

Allogeneic islet transplantation offers a promising therapy for patients with type 1 diabetes, this method is still inefficient because of considerable islet loss shortly after transplantation [1, 2]. The main islet vasculature is damaged or destroyed. Mass transport of relevant nutrients mainly depends on passive diffusion, which adversely affects the viability of isolated human islets [3]. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

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