Assessment of β-Cell Replication in Isolated Rat Islets of Langerhans

  • Louise T. DalgaardEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2029)


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.

Key words

Islet of Langerhans β-Cell Proliferation Mitosis Ethynyldeoxyuridine (EdU) Glucose Insulin Hoechst 33342 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Science and EnvironmentRoskilde UniversityRoskildeDenmark

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