Abstract
Pancreatic β-cells in the islets of Langerhans secrete insulin in response to the rise in glucose levels following food intake. The hypoglycemic action of insulin applies a strong evolutionary brake on β-cell division. However, under some conditions β-cells can be stimulated to enter cell cycle progression and divide, for example following exposure to increased glucose levels or during pregnancy. Here, a protocol is described for the isolation of rat adult islets of Langerhans, followed by culture of intact islets in Matrigel and measurement of β-cell replication by the incorporation of ethynyldeoxyuridine (EdU). EdU positive cells are revealed by a click reaction, nuclei are visualized using a DNA-binding fluorophore (Hoechst 33342), and β-cells are identified using immunofluorescence detection.
Originalsprog | Engelsk |
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Tidsskrift | Methods in Molecular Biology |
Vol/bind | 2029 |
Sider (fra-til) | 25-35 |
Antal sider | 11 |
ISSN | 1064-3745 |
DOI | |
Status | Udgivet - 2019 |