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

Louise Torp Dalgaard

doi:10.1007/978-1-4939-9631-5_3

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

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

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
Tidsskrift	Methods in Molecular Biology
Vol/bind	2029
Sider (fra-til)	25-35
Antal sider	11
ISSN	1064-3745
DOI	https://doi.org/10.1007/978-1-4939-9631-5_3
Status	Udgivet - 2019

Bibliografisk note

Important note from the Publisher regarding the attached version of the article: “This is a post-peer-review, pre-copyedit version of an article published in Methods in Molecular Biology. The final authenticated version is available online at: http://dx.doi.org/10.1007/978-1-4939-9631-5_3”.

Link til dokument

10.1007/978-1-4939-9631-5_3

Assessment of glucose-induced beta cell proliferationAccepteret manuskript, 299 KB

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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.",

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AB - 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.

KW - Ethynyldeoxyuridine (EdU)

KW - Glucose

KW - Hoechst 33342

KW - Insulin

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KW - Mitosis

KW - Proliferation

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