In vitro formation of β cell pseudoislets using islet-derived endothelial cells

• Published in: PloS one 2013-08, Vol.8 (8), p.e72260-e72260
• Main Authors: Spelios, Michael G, Kenna, Lauren A, Wall, Bonnie, Akirav, Eitan M
• Format: Article
• Language: English
• Subjects: Animals; Antibiotics; Apoptosis; Biology; Cell culture; Cell Culture Techniques; Cell Differentiation; Cell Line, Tumor; Collagen Type IV - metabolism; Configurations; Endothelial cells; Endothelial Cells - physiology; Extracellular matrix; Floating structures; Gene expression; Glucose; Immunoglobulins; Insulin; Insulin secretion; Insulin-Secreting Cells - physiology; Islets of Langerhans - cytology; Laminin - metabolism; Mechanical properties; Mice; Neuroendocrine tumors; Proteins; Rodents; Secretion; Transplants & implants
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0072260

β cell pseudoislets (PIs) are used for the in vitro study of β-cells in a three-dimensional (3-D) configuration. Current methods of PI induction require unique culture conditions and extensive mechanical manipulations. Here we report a novel co-culture system consisting of high passage β-cells and islet-derived endothelial cells (iECs) that results in a rapid and spontaneous formation of free-floating PIs. PI structures were formed as early as 72 h following co-culture setup and were preserved for more than 14 d. These PIs, composed solely of β-cells, were similar in size to that of native islets and showed an increased percentage of proinsulin-positive cells, increased insulin gene expression in response to glucose stimulation, and restored glucose-stimulated insulin secretion when compared to β-cells cultured as monolayers. Key extracellular matrix proteins that were absent in β-cells cultured alone were deposited by iECs on PIs and were found in and around the PIs. iEC-induced PIs are a readily available tool for examining β cell function in a native 3-D configuration and can be used for examining β-cell/iEC interactions in vitro.