title: Controlled aggregation of primary human pancreatic islet cells leads to glucose-responsive pseudoislets comparable to native islets.
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.
authors.affiliation: Department of Developmental Bioengineering, University of Twente, Enschede, The Netherlands.
authors.forename: Janneke
authors.initials: J
authors.lastname: Hilderink
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Siebe
authors.initials: S
authors.lastname: Spijker
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Françoise
authors.initials: F
authors.lastname: Carlotti
authors.affiliation: Department of Developmental Bioengineering, University of Twente, Enschede, The Netherlands.
authors.forename: Lydia
authors.initials: L
authors.lastname: Lange
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Marten
authors.initials: M
authors.lastname: Engelse
authors.affiliation: Department of Tissue Regeneration, University of Twente, Enschede, The Netherlands.
authors.forename: Clemens
authors.initials: C
authors.lastname: van Blitterswijk
authors.affiliation: Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands.|Hubrecht Institute, Utrecht, The Netherlands.
authors.forename: Eelco
authors.initials: E
authors.lastname: de Koning
authors.affiliation: Department of Developmental Bioengineering, University of Twente, Enschede, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Karperien
authors.affiliation: Department of Developmental Bioengineering, University of Twente, Enschede, The Netherlands.
authors.forename: Aart
authors.identifier: http://orcid.org/0000-0002-2940-5699
authors.initials: A
authors.lastname: van Apeldoorn
mesh_terms: D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D045744:Cell Line, Tumor; D048429:Cell Size; D002470:Cell Survival; D002478:Cells, Cultured; D005260:Female; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007340:Insulinoma / Q000473:pathology; D007515:Islets of Langerhans / Q000166:cytology*; D008297:Male; D016513:Mice, SCID; D008875:Middle Aged; D015203:Reproducibility of Results
chemical_list: D005947:Glucose
keywords: beta cells; islets; pseudoislets bioengineering; type 1 diabetes