CFSE staining for T cell Functional Assays

March 05, 2015

University of Nebraska Medical Center
Pharmacology and Experimental Neuroscience
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Life Technologies

Product Name:

CellTrace™ CFSE Cell Proliferation Kit

Catalog Number:

C34554

The general aim of my research is to address the role of T cells in neuronal degeneration in Parkinson's disease. My specific goals for the project in which this kit was used was to address the ability of regulatory T cells isolated from MPTP-intoxicated mice to suppress the proliferation of effector T cell populations isolated from the same mice. I chose this kit because it allows for fluorescent labeling of cells to look at proliferation.

Experimental Design and Results Summary

Application

Fluorescent labeling of cells

Starting Material

CD4+CD25- T cells

Protocol Overview

I used this kit to fluorescently label effector T cells (CD4+CD25-) isolated from spleens and lymph nodes of MPTP-intoxicated mice in order to look at cell proliferation.

Tips

You will definitely need to triturate each new lot! The optimized concentration of CFSE will change with each lot. It can also be harsh on the cells if you leave them in the staining buffer/CFSE dye for too long.

Results Summary

I am able to successfully detect cell proliferation and generations using the CFSE fluorescent dye. The cell viability following labeling is also good. The cells continue to survive in culture and divide well. They also look healthy. However, sometimes I do get uneven labeling resulting in fluorescent peaks.

Additional Notes

None

Image Gallery

Summary

The Good

Cells are viable following labeling, inexpensive, fairly easy to use

The Bad

Have to optimize each new lot, can be toxic to cells

The Bottom Line

I use this kit all the time and it works well for my experiments. I can easily stain cells to look for proliferative capabilities. I recommend the product; however, getting the staining protocol optimized took me some time and practice.

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