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-1dim Ly-6G– cells can be further subdivided according to the expression of Ly-6C into Gr-1dim Ly-6Chigh Ly-6G– and Gr-1dim Ly-6Clow Ly-6G– cells. The proportion of Ly-6Chigh and Ly-6Clow cells varies among tumor models.
Detailed separation procedure
The Myeloid-Derived Suppressor Cell Isolation Kit has been developed for the isolation of Gr-1high Ly-6G+ and Gr-1dim Ly-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-1high Ly-6G+ cells. The unlabeled cells flow through; this cell fraction contains Gr-1dim Ly-6G– myeloid cells. The magnetically labeled Gr-1high Ly-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-1dim Ly-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 (depletion): LS or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
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