Isolation Of Murine CD4+ Cells Using Magnetic Bead Isolation

March 23, 2017

Albert Einstein College of Medicine
Pathology
PhD Candidate

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Company:

Thermo Fisher Scientific

Product Name:

Dynabeads® Mouse CD4

Catalog Number:

11445D

An ongoing study in our lab is examining how a protein of interest functions in CD4+ T cells during inflammation in the CNS. To isolate CD4+ T cells for in vitro studies, we routinely utilize the Dynabeads for mouse CD4 (L3T4) and isolate using magnetic bead separation.

Experimental Design and Results Summary

Application

Cell isolation

Starting Material

Mouse lymph nodes and spleen

Protocol Overview

Isolate lymph nodes and spleen, mechanically grind tissues in 40um filter immersed in cell culture media, wash CD4 magnetic beads in media, add to homogenized tissue, incubate for 35 min at 4*C, apply tube to magnet, aspirate media and wash cells, aspirate media and resuspend cells in media, add DetachaBead reagent and incubate at RT for 45 min to cells to release cells from magnetic beads

Tips

Be sure to make sure the beads are all in suspension before beginning. The beads tend to settle and stick to the bottom of the bottle.

Results Summary

When isolating cells from lymph nodes and spleen, a high yield of 1-2x10^7 cells is achieved with high purity and high viability.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

High cell yield and viability for downstream applications

The Bad

Beads are expensive and the DetachaBead reagent is needed to detach the cells from the Dynabeads, can be somewhat time consuming

The Bottom Line

Great way to get pure populations of CD4+ cells with high viability for experiments.

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