Anti-Human CD8 Microbeads - A Consistent and Reliable Method to Positively Select CD8+ T Cells from PBMC

May 12, 2016

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

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Quality of Results

Ease-of-Optimization

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Company:

Miltenyi

Product Name:

CD8 Microbeads, human

Catalog Number:

130-045-201

Our lab routinely works with purified immune cell subsets from human peripheral blood mononuclear cells (PBMC). In order to obtain highly pure CD8+ T cells, as well as other immune cell types, we routinely employ magnetic separation with Miltenyi products for positive selection because of their ease of use and minimal optimization. We require highly pure CD8+ T cells due to the nature of our downstream analysis, and this product helps us achieve this goal.

Experimental Design and Results Summary

Application

Positive selection of CD8+ 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. CD8+ microbeads were used to isolate CD8+ cells from PBMC by positive selection according to the protocol defined in the product datasheet when using an autoMACS Pro Separator. This separation was performed in combination with the positive selection of other cell types.

Tips

The protocol provided by the manufacturer is very detailed, thus little optimization was required. Some points for consideration would be to increase the incubation time of CD8 microbeads with your cells to increase the yield; however, this may lead to an unwanted decrease in purity. Ultimately, if you are using an autoMACS Pro Separator for your magnetic separation, you may want to try different separation programs to optimize and balance yield vs purity instead of varying the incubation times and bead to cell incubation ratios.

Results Summary

A highly pure population of CD8+ T cells was obtained.

Additional Notes

None

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Summary

The Good

Minimal optimization. Comes ready to use. Well established protocols. Can be combined with other positive-selection products so that multiple cell types can be obtained from a single biological sample if so desired. Purity of isolated cells was excellent.

The Bad

Knowledge of/access to flow cytometric services is required to perform validation steps after using these beads to isolate CD8+ T cells. Depending on your downstream experiments, you may require untouched CD8 T cells, in which case you should consider employing a negative selection technique as an alternative to this product. Product is only available in one size.

The Bottom Line

If you want to separate CD8+ T cells from a heterogeneous cell population such as PBMC, and cell purity is a priority in you downstream experiments, this product is highly recommended.

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