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

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

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.
OriginalsprogEngelsk
TidsskriftMethods in Molecular Biology
Vol/bind2029
Sider (fra-til)25-35
Antal sider11
ISSN1064-3745
DOI
StatusUdgivet - 2019

Citer dette

@article{c5342909bc1f418592bdf7fbc19e4389,
title = "Assessment of β-Cell Replication in Isolated Rat Islets of Langerhans",
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.",
author = "Dalgaard, {Louise Torp}",
year = "2019",
doi = "10.1007/978-1-4939-9631-5_3",
language = "English",
volume = "2029",
pages = "25--35",
journal = "Methods in Molecular Biology",
issn = "1064-3745",
publisher = "Humana Press",

}

Assessment of β-Cell Replication in Isolated Rat Islets of Langerhans. / Dalgaard, Louise Torp.

I: Methods in Molecular Biology, Bind 2029, 2019, s. 25-35.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Assessment of β-Cell Replication in Isolated Rat Islets of Langerhans

AU - Dalgaard, Louise Torp

PY - 2019

Y1 - 2019

N2 - 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.

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.

U2 - 10.1007/978-1-4939-9631-5_3

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

M3 - Journal article

VL - 2029

SP - 25

EP - 35

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -