Human pancreatic islet-derived extracellular vesicles modulate insulin expression in 3D-differentiating iPSC clusters

• Published in: PloS one Vol. 12; no. 11; p. e0187665
• Main Authors: Ribeiro, Diana, Andersson, Eva-Marie, Heath, Nikki, Persson-Kry, Anette, Collins, Richard, Hicks, Ryan, Dekker, Niek, Forslöw, Anna
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 08-11-2017; Public Library of Science (PLoS)
• Subjects: Antigens, CD - metabolism; Bioengineering; Biology and Life Sciences; Biomarkers - metabolism; Bone marrow; CD63 antigen; CD81 antigen; Cell Aggregation - drug effects; Cell differentiation; Cell Differentiation - drug effects; Clusters; Collagen; Collagen - pharmacology; Crosstalk; Differentiation (biology); Enzyme-Linked Immunosorbent Assay; Extracellular vesicles; Extracellular Vesicles - drug effects; Extracellular Vesicles - metabolism; Extracellular Vesicles - ultrastructure; Gene expression; Genetic aspects; Glucagon; Hormones; Humans; Hydrogel, Polyethylene Glycol Dimethacrylate - pharmacology; Hydrogels; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - ultrastructure; Insulin; Insulin - metabolism; Insulin resistance; Intracellular; Intracellular levels; Islets of Langerhans - cytology; Laboratories; Markers; Medicine and Health Sciences; Metabolism; Metabolites; MicroRNAs; Pancreas; Physical Sciences; Physiological aspects; Pluripotency; Proteins; Rodents; Stem cells; Suspensions; Time Factors; Tissue engineering; Transcription factors; Vesicles; Sweden
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0187665

It has been suggested that extracellular vesicles (EVs) can mediate crosstalk between hormones and metabolites within pancreatic tissue. However, the possible effect of pancreatic EVs on stem cell differentiation into pancreatic lineages remains unknown. Herein, human islet-derived EVs (h-Islet-EVs) were isolated, characterized and subsequently added to human induced pluripotent stem cell (iPSC) clusters during pancreatic differentiation. The h-islet-EVs had a mean size of 117±7 nm and showed positive expression of CD63 and CD81 EV markers as measured by ELISA. The presence of key pancreatic transcription factor mRNA, such as NGN3, MAFA and PDX1, and pancreatic hormone proteins such as C-peptide and glucagon, were confirmed in h-Islet-EVs. iPSC clusters were differentiated in suspension and at the end stages of the differentiation protocol, the mRNA expression of the main pancreatic transcription factors and pancreatic hormones was increased. H-Islet-EVs were supplemented to the iPSC clusters in the later stages of differentiation. It was observed that h-Islet-EVs were able to up-regulate the intracellular levels of C-peptide in iPSC clusters in a concentration-dependent manner. The effect of h-Islet-EVs on the differentiation of iPSC clusters cultured in 3D-collagen hydrogels was also assessed. Although increased mRNA expression for pancreatic markers was observed when culturing the iPSC clusters in 3D-collagen hydrogels, delivery of EVs did not affect the insulin or C-peptide intracellular content. Our results provide new information on the role of h-Islet-EVs in the regulation of insulin expression in differentiating iPSC clusters, and are highly relevant for pancreatic tissue engineering applications.