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Murine Pancreatic Islet Isolation

作者： Gregory L. Szot1, Pavel Koudria1, Jeffrey A. Bluestone1 1Diabetes Center,University of California, San Francisco - UCSF

简介：

Overview

This video demonstrates the process of isolating mouse islets from the pancreas of a C3H donor mouse. The procedure includes collagenase injection, pancreas removal, and islet purification for transplantation into a diabetic NOD mouse.

Key Study Components

Area of Science

Neuroscience
Endocrinology
Transplantation Biology

Background

Islet transplantation is a potential treatment for diabetes.
Isolation of islets requires specific enzymatic digestion.
Collagenase is commonly used for islet isolation.
Mouse models are frequently used in diabetes research.

Purpose of Study

To demonstrate a reliable method for isolating mouse islets.
To prepare islets for transplantation into diabetic recipients.
To enhance understanding of islet function and viability post-isolation.

Methods Used

Injection of collagenase solution into the pancreas.
Removal and digestion of the pancreas.
Purification of islets using a Ficoll gradient.
Handpicking of islets for transplantation.

Main Results

Successful isolation of islets from the pancreas.
Islets prepared for transplantation showed viability.
Method demonstrated effectiveness for future studies.
Potential for improving diabetes treatment through transplantation.

Conclusions

The described method is efficient for isolating mouse islets.
Islet transplantation can be a viable option for diabetic models.
Further research can build on this isolation technique.

Frequently Asked Questions

What is the significance of isolating mouse islets?

Isolating mouse islets is crucial for studying diabetes and testing transplantation methods.

What role does collagenase play in this procedure?

Collagenase is used to digest the pancreas and release the islets for isolation.

Why use a C3H mouse model?

C3H mice are known for their reliable islet yield and are commonly used in research.

What is the purpose of transplanting islets into NOD mice?

NOD mice are a model for diabetes, allowing researchers to study islet function in a diabetic context.

How are the islets purified after digestion?

Islets are purified using a Ficoll gradient to separate them from other pancreatic tissue.

What are the potential outcomes of this research?

The research may lead to improved techniques for islet transplantation and better diabetes treatments.

Hi, I am Greg Zott. I'm with the UCSF Diabetes Center in the Bluestone Laboratory, and today I'm gonna show you the process for isolating mouse eyelets. The procedure involves the injection of a collagenase solution into the pancreas of a C3 H donor mouse, where we will then remove that pancreas, digest it away, purify the eyelets on a fial gradient, and then handpick them for ultimate transplant into the kidney capsule of a diabetic NOD mouse, Which will be our recipient.

Before we start this procedure, we diluted our ATE solution in a modified Hanks buffer and we have adjusted the concentration specific for the strain of mouse that we're using the C3 H.We found digests better at a 0.8 mgs per mil collagenase solution, and we will use that to inject into the common bile duct to distend the pancreas.Performing. The procedure today Will be Pavo codre of the diabetes center. He will inject, open up the mouse, expose the the intestines, and then inject into the common bile duct.

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