Human EndoC-βH1 β-cells form pseudoislets with improved glucose sensitivity and enhanced GLP-1 signaling in the presence of islet-derived endothelial cells
| Autor: | Eitan M. Akirav, Regine C Tipon, Lauren A Afinowicz, Michael G. Spelios |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
endocrine system medicine.medical_specialty Physiology Endocrinology Diabetes and Metabolism 030209 endocrinology & metabolism 03 medical and health sciences Islets of Langerhans 0302 clinical medicine Glucagon-Like Peptide 1 Physiology (medical) Internal medicine Insulin-Secreting Cells Glucose Intolerance medicine Humans Insulin Insulinoma Cells Cultured geography geography.geographical_feature_category Glucose sensitivity Chemistry Endothelial Cells Islet medicine.disease Glucagon-like peptide-1 Coculture Techniques Endothelial stem cell 030104 developmental biology Endocrinology Glucose Beta cell Insulin Resistance Signal Transduction |
| Zdroj: | American journal of physiology. Endocrinology and metabolism. 314(5) |
| ISSN: | 1522-1555 |
| Popis: | Three-dimensional (3D) pseudoislets (PIs) can be used for the study of insulin-producing β-cells in free-floating islet-like structures similar to that of primary islets. Previously, we demonstrated the ability of islet-derived endothelial cells (iECs) to induce PIs using murine insulinomas, where PI formation enhanced insulin production and glucose responsiveness. In this report, we examined the ability of iECs to spontaneously induce the formation of free-floating 3D PIs using the EndoC-βH1 human β-cell line murine MS1 iEC. Within 14 days, the coculturing of both cell types produced fully humanized EndoC-βH1 PIs with little to no contaminating murine iECs. The size and shape of these PIs were similar to primary human islets. iEC-induced PIs demonstrated reduced dysregulated insulin release under low glucose levels and higher insulin secretion in response to high glucose and exendin-4 [a glucagon-like peptide-1 (GLP-1) analog] compared with monolayer cells cultured alone. Interestingly, iEC-PIs were also better at glucose sensing in the presence of extendin-4 compared with PIs generated on a low-adhesion surface plate in the absence of iECs and showed an overall improvement in cell viability. iEC-induced PIs exhibited increased expression of key genes involved in glucose transport, glucose sensing, β-cell differentiation, and insulin processing, with a concomitant decrease in glucagon mRNA expression. The enhanced responsiveness to exendin-4 was associated with increased protein expression of GLP-1 receptor and phosphokinase A. This rapid coculture system provides an unlimited number of human PIs with improved insulin secretion and GLP-1 responsiveness for the study of β-cell biology. |
| Databáze: | OpenAIRE |
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