Myeloid-Derived Suppressor Cell Isolation Kit ( #130-094-538 )

Myeloid-Derived Suppressor Cell Isolation Kit

Overview

The Myeloid-Derived Suppressor Cell Isolation Kit has been developed for the isolation of Gr-1^highLy-6G⁺ and Gr-1^dimLy-6G^– myeloid cells from lymphoid tissue.

Details

Background information
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells, characterized as CD11b⁺Gr-1⁺, with a remarkable capacity to induce T cell dysfunction. They arise in various pathological conditions (cancer, infection, autoimmune diseases), where the differentiation of immature myeloid progenitor cells into granulocytes, macrophages, or dendritic cells is partially blocked.
Gr-1^dimLy-6G^– cells can be further subdivided according to the expression of Ly-6C into Gr-1^dimLy-6C^highLy-6G^– and Gr-1^dimLy-6C^lowLy-6G^– cells. The proportion of Ly-6C^high and Ly-6C^low cells varies among tumor models.

Detailed separation procedure
The Myeloid-Derived Suppressor Cell Isolation Kit has been developed for the isolation of Gr-1^highLy-6G⁺ and Gr-1^dimLy-6G^– myeloid cells.

First, the cell suspension is incubated with an Anti-Ly-6G-Biotin antibody and Anti-Biotin MicroBeads. The cells are subsequently applied to a MACS Column, which retains the magnetically labeled Gr-1^highLy-6G⁺ cells. The unlabeled cells flow through; this cell fraction contains Gr-1^dimLy-6G^– myeloid cells. The magnetically labeled Gr-1^highLy-6G⁺ cells can be eluted as the positively selected cell fraction and are further purified by applying them to a second MACS Column.

Second, the pre-enriched Gr-1^dimLy-6G^– subpopulation is further purified. To this end, the cells are incubated with an Anti-Gr-1-Biotin antibody and Streptavidin MicroBeads, and isolated by positive selection over two MACS Columns.

Downstream applications
MDSCs isolated with this kit can be used for functional analyses, such as T cell activation and effector function in experimental animal models. Isolated MDSCs are also suitable for, e.g., phenotypical analysis or downstream gene expression profiling.

Columns

For the first magnetic separation: LS or autoMACS Columns; for the second magnetic separation: MS or autoMACS Columns.

Figure 1

Myeloid-derived suppressor cells were isolated from a spleen of a BALB/c mouse using the Myeloid-Derived Suppressor Cell Isolation Kit, two LS Columns, two MS Columns, and an appropriate MACS Separator. Cells were fluorescently stained with CD11b-APC and Anti-Gr-1-PE and analyzed by flow cytometry. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Spleen cells before separation

Enriched Gr-1^highLy-6G⁺ cells

Enriched Gr-1<sup>high</sup>Ly-6G<sup>+</sup> cells

Enriched Gr-1^dimLy-6G^– cells

Enriched Gr-1<sup>dim</sup>Ly-6G<sup>–</sup> cells