Microbiology JoVE (Open Access) Citable · DOI

Mouse Islet of Langerhans Isolation using a Combination of Purified Collagenase and Neutral Protease

Name: Mouse Islet of Langerhans Isolation using a Combination of Purified Collagenase and Neutral Protease
Uploaded: 2026-06-17
Duration: 6 min 59 s
Description: The interrogation of beta cell gene expression and function in vitro has squarely shifted over the years from the study of rodent tumorigenic cell lines ...

DOI: 10.3791/4137-v

What you'll learn

✓ Perform pancreatic ductal cannulation and enzymatic perfusion in mice
✓ Isolate intact islets of Langerhans using collagenase and neutral protease
✓ Dissociate pancreatic tissue and recover functional islet preparations

Protocol

A detailed description of mouse islet isolation is described using the technique of in situ pancreatic ductal cannulation and perfusion of a combination of purified collagenase and neutral protease.

Difficulty

advanced

Total time

~45–60 min per mouse

Model organism

Mouse (strain unspecified)

Biosafety

BSL-1

Steps

Inflate pancreas with collagenase-protease mixture

Cannulate the pancreatic duct and perfuse with a combination of purified collagenase and neutral protease to enzymatically digest the exocrine tissue and loosen islets from surrounding acinar cells.

▶ 01:20

Dissociate pancreatic tissue into single islets

Mechanically dissociate the enzyme-treated pancreatic tissue through gentle pipetting and trituration to separate individual islets of Langerhans from remaining acinar debris.

▶ 03:58

Collect and assess isolated islet preparation

Recover the isolated islets through centrifugation or density-based separation and evaluate islet yield and morphology under microscopy to confirm successful isolation.

▶ 05:49

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