Abstract
| Original language | English |
|---|---|
| Journal | Methods in Molecular Biology |
| Volume | 2029 |
| Pages (from-to) | 25-35 |
| Number of pages | 11 |
| ISSN | 1064-3745 |
| DOIs | |
| Publication status | Published - 2019 |
Cite this
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Assessment of β-Cell Replication in Isolated Rat Islets of Langerhans. / Dalgaard, Louise Torp.
In: Methods in Molecular Biology, Vol. 2029, 2019, p. 25-35.Research output: Contribution to journal › Journal article › Research › peer-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 -