Artikel ; Online: Hypoxia induction in cultured pancreatic islets enhances endothelial cell morphology and survival while maintaining beta-cell function.
PloS one
2019 Band 14, Heft 10, Seite(n) e0222424
Abstract: Background: Pancreatic islets are heavily vascularized in vivo yet lose this vasculature after only a few days in culture. Determining how to maintain islet vascularity in culture could lead to better outcomes in transplanting this tissue for the ... ...
| Abstract | Background: Pancreatic islets are heavily vascularized in vivo yet lose this vasculature after only a few days in culture. Determining how to maintain islet vascularity in culture could lead to better outcomes in transplanting this tissue for the treatment of type 1 diabetes as well as provide insight into the complex communication between beta-cells and endothelial cells (ECs). We previously showed that islet ECs die in part due to limited diffusion of serum albumin into the tissue. We now aim to determine the impact of hypoxia on islet vascularization. Methods: We induced hypoxia in cultured mouse islets using the hypoxia mimetic cobalt chloride (100 μM CoCl2). We measured the impact on islet metabolism (two-photon NAD(P)H and Rh123 imaging) and function (insulin secretion and survival). We also measured the impact on hypoxia related transcripts (HIF-1α, VEGF-A, PDK-1, LDHA, COX4) and confirmed increased VEGF-A expression and secretion. Finally, we measured the vascularization of islets in static and flowing culture using PECAM-1 immunofluorescence. Results: CoCl2 did not induce significant changes in beta cell metabolism (NAD(P)H and Rh123), insulin secretion, and survival. Consistent with hypoxia induction, CoCl2 stimulated HIF-1α, PDK-1, and LDHA transcripts and also stimulated VEGF expression and secretion. We observed a modest switch to the less oxidative isoform of COX4 (isoform 1 to 2) and this switch was noted in the glucose-stimulated cytoplasmic NAD(P)H responses. EC morphology and survival were greater in CoCl2 treated islets compared to exogenous VEGF-A in both static (dish) and microfluidic flow culture. Conclusions: Hypoxia induction using CoCl2 had a positive effect on islet EC morphology and survival with limited impact on beta-cell metabolism, function, and survival. The EC response appears to be due to endogenous production and secretion of angiogenic factors (e.g. VEGF-A), and mechanistically independent from survival induced by serum albumin. |
|---|---|
| Mesh-Begriff(e) | Animals ; Cell Communication/drug effects ; Cell Communication/genetics ; Cell Hypoxia/drug effects ; Cell Hypoxia/genetics ; Cobalt/pharmacology ; Electron Transport Complex IV/genetics ; Endothelial Cells/cytology ; Endothelial Cells/drug effects ; Endothelial Cells/metabolism ; Gene Expression Regulation/genetics ; Glucose/metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics ; Insulin/genetics ; Insulin/metabolism ; Insulin Secretion/genetics ; Insulin-Secreting Cells/cytology ; Insulin-Secreting Cells/drug effects ; Insulin-Secreting Cells/metabolism ; Islets of Langerhans/cytology ; Islets of Langerhans/drug effects ; Islets of Langerhans/metabolism ; Mice ; Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase/genetics ; Vascular Endothelial Growth Factor A/genetics |
| Chemische Substanzen | Hif1a protein, mouse ; Hypoxia-Inducible Factor 1, alpha Subunit ; Insulin ; Pdk1 protein, mouse ; Pyruvate Dehydrogenase (Acetyl-Transferring) Kinase ; Vascular Endothelial Growth Factor A ; vascular endothelial growth factor A, mouse ; Cobalt (3G0H8C9362) ; Cox4i1 protein, mouse (EC 1.9.3.1) ; Electron Transport Complex IV (EC 1.9.3.1) ; cobaltous chloride (EVS87XF13W) ; Glucose (IY9XDZ35W2) |
| Sprache | Englisch |
| Erscheinungsdatum | 2019-10-10 |
| Erscheinungsland | United States |
| Dokumenttyp | Journal Article ; Research Support, Non-U.S. Gov't |
| ISSN | 1932-6203 |
| ISSN (online) | 1932-6203 |
| DOI | 10.1371/journal.pone.0222424 |
| Datenquelle | MEDical Literature Analysis and Retrieval System OnLINE |
Zusatzmaterialien
Kategorien
Über subito bestellen
Dieser Service ist kostenpflichtig (siehe Lieferbedingungen von subito). Bestellungen, die einen Artikel nebst Supplementary Material umfassen, werden grundsätzlich wie mehrfache Bestellungen bearbeitet. Gebühren fallen in diesen Fällen für jede einzelne Bestellung an.
Fernleihe an ZB MED
Sie können sich den gewünschten Titel als lokale Nutzerin oder lokaler Nutzer von ZB MED direkt an den Standort Köln schicken lassen.