Miltenyi CD4 Microbeads, Human - Reliable Method for the Magnetic Positive Selection of CD4+ T Cells from Human PBMC

May 16, 2016

Memorial University of Newfoundland
Division of BioMedical Sciences, Faculty of Medicine
Research Assistant

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Company:

Miltenyi

Product Name:

CD4 Microbeads, human

Catalog Number:

130-045-101

Several of our in vitro experiments require purified primary CD4+ T cells . We have worked with total PBMC in the past, but found that using purified CD4+ T cells gave more consistent results. After comparing different cell separation technologies, we settled upon CD4+ Microbeads from Miltenyi given their consistency, quick protocol, and purity of the isolated cells.

Experimental Design and Results Summary

Application

Positive selection of CD4+ cells from human PBMC using an autoMACS Pro Separator

Starting Material

Human Peripheral blood mononuclear cells (PBMC) isolated from whole blood

Protocol Overview

Human PBMCs were isolated from whole blood by standard Ficoll density gradient separation. CD4+ microbeads were used to isolate CD4+ cells from PBMC by positive selection according to the exact protocol outlined in the product datasheet when using an autoMACS Pro Separator. This separation was performed in combination with the positive selection of other cell types, with this separation occurring at the end.

Tips

The ability of additional cell populations to express CD4 other than CD4 T cells (monocytes for example), should be taken into consideration when using this product for separation. Depending on the requirements of your downstream experiments concerning the purity of the separated cells, you make want to remove monocytes before CD4 separation by either adherence or CD14 positive selection. We observed some monocytes still remaining in our CD4 positively-selected cells even after prior removal of monocytes by CD14 positive selection, but the amount of contamination as minimal.

Results Summary

A relatively pure population of CD4+ cells was obtained.

Additional Notes

None

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Summary

The Good

Relatively simple protocol. Quick and straightforward.

The Bad

Additional flow cytometric analysis required for validation of the technique. Product is only available in one size.

The Bottom Line

This product should be seriously considered for researchers who wish to purify CD4+ T cells from biological samples such as human PBMC.

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