Bioluminescent Monitoring of Graft Survival in an Adoptive Transfer Model of Autoimmune Diabetes in Mice

Taylor Stewart; Kevin Bode; Stephan Kissler; Peng Yi

doi:10.3791/64836

Bioluminescent Monitoring of Graft Survival in an Adoptive Transfer Model of Autoimmune Diabetes ...

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Bioluminescent Monitoring of Graft Survival in an Adoptive Transfer Model of Autoimmune Diabetes in Mice

Full Text

2,357 Views

10:03 min

November 18, 2022

DOI: 10.3791/64836-v

Taylor Stewart¹, Kevin Bode², Stephan Kissler², Peng Yi¹

¹Section for Islet Cell and Regenerative Biology, Joslin Diabetes Center,Harvard Medical School, ²Section for Immunology, Joslin Diabetes Center,Harvard Medical School

Overview

This protocol presents a minimally invasive and cost-effective method for transplanting and imaging NIT-1 cells in non-obese diabetic (NOD)-severe combined immunodeficient mice. The approach aims to enhance the understanding of autoimmune responses and therapeutic targets for beta cell replacement in type one diabetes.

Key Study Components

Research Area

Type one diabetes
Autoimmune disease mechanisms
Cell transplantation techniques

Background

Immune response in diabetes
Need for beta cell replacement therapies
Challenges in current methodologies

Methods Used

Cell transplant techniques
Non-obese diabetic (NOD) mice model
Imaging methods for tracking cell behavior

Main Results

Successful cell transplant in mouse model
Demonstrated low invasiveness and ease of execution
Potential for identifying therapeutic targets

Conclusions

The study illustrates a practical method for studying autoimmune attacks against beta cells.
This technique contributes to future research in diabetes treatment strategies.

Frequently Asked Questions

What is the purpose of this protocol?

To study mechanisms of protection against autoimmune attacks and target therapies for diabetes.

What are NIT-1 cells?

NIT-1 cells are insulin-producing cells used in diabetes research.

Who can perform the tail vein injection?

It is recommended to have an expert demonstrate the method for beginners.

What is the main advantage of this transplantation method?

The method is cost-effective, simple, and minimally invasive.

How are the cells prepared before transplantation?

Cells are purified from the spleen and resuspended in PBS before injection.

What imaging technique is used post-transplant?

D-luciferin is administered for bioluminescent imaging.

What does the study contribute to diabetes research?

It provides insights into transplantation techniques and potential therapeutic targets.

This protocol describes a straightforward and minimally invasive method for transplanting and imaging NIT-1 cells in non-obese diabetic (NOD)-severe combined immunodeficient mice challenged with splenocytes purified from spontaneously diabetic NOD mice.

This protocol can help to understand the mechanisms of protection against autoimmune attacks and identify therapeutic targets for beta cell replacement in type one diabetes. The main advantage of this technique is the relatively low cost, the simplicity of the cell culture, transplant, and imaging methods, as well as minimal invasiveness of the mouse techniques. Individuals who have never performed a tail vein injection before may struggle to insert the needle into the vein.

Ask for a demonstration by an expert before your first injection. To begin, take the anesthetized mouse and remove hair from its dorsal side using an electric shaver with a guard, exposing approximately one inch by two inches of the skin. Return the shaved mouse to the knockdown chamber.

View the full transcript and gain access to thousands of scientific videos