Myeloid-Derived Suppressor Cell Isolation Kit, mouse

Name: Myeloid-Derived Suppressor Cell Isolation Kit, mouse
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Myeloid-Derived Suppressor Cell Isolation Kit, mouse

The Myeloid-Derived Suppressor Cell Isolation Kit has been developed for the isolation of Gr-1

^high

Ly-6G

⁺

and Gr-1

^dim

Ly-6G

^–

myeloid cells from lymphoid tissue.

This Kit works ideally for spleen and tumor tissues.

Product data and images for Myeloid-Derived Suppressor Cell Isolation Kit, mouse

Figures

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 ^high Ly-6G ⁺ cells
View details Myeloid-Derived Suppressor Cell Isolation Kit, mouse 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.	View details Myeloid-Derived Suppressor Cell Isolation Kit, mouse 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.
Enriched Gr-1 ^dim Ly-6G ^– cells
View details Myeloid-Derived Suppressor Cell Isolation Kit, mouse 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.

Specifications for Myeloid-Derived Suppressor Cell Isolation Kit, mouse

Overview

The Myeloid-Derived Suppressor Cell Isolation Kit has been developed for the isolation of Gr-1

^high

Ly-6G

⁺

and Gr-1

^dim

Ly-6G

^–

myeloid cells from lymphoid tissue.

This Kit works ideally for spleen and tumor tissues.

Detailed product information

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

^dim

Ly-6G

^–

cells can be further subdivided according to the expression of Ly-6C into Gr-1

^dim

Ly-6C

^high

Ly-6G

^–

and Gr-1

^dim

Ly-6C

^low

Ly-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

^high

Ly-6G

⁺

and Gr-1

^dim

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-1

^high

Ly-6G

⁺

cells. The unlabeled cells flow through; this cell fraction contains Gr-1

^dim

Ly-6G

^–

myeloid cells. The magnetically labeled Gr-1

^high

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-1

^dim

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.

Resources for Myeloid-Derived Suppressor Cell Isolation Kit, mouse

Documents and Protocols

Brochures/posters

Isolation and analysis of murine myeloid cells from lymphoid tissues and tumor

Brochures/posters

All you need for myeloid cell research - Quick guide

References for Myeloid-Derived Suppressor Cell Isolation Kit, mouse

Publications

Serafini, R. et al. (2004) Derangement of immune responses by myeloid suppressor cells. Cancer Immunol. Immunother. 53: 64-72
Youn, J. et al. (2008) Subsets of myeloid-derived suppressor cells in tumor-bearing mice. J. Immunol. 181: 5791-5802
Ostrand-Rosenberg, S. and Sinha, P. (2009) Myeloid-derived suppressor cells: linking inflammation and cancer. J. Immunol. 182: 4499-4506
Gabrilovich, D. I. and Nagaraj, S. (2009) Myeloid-derived suppressor cells as regulators of the immune system. Nat. Rev. Immunol. 9: 162-174
Bronte, V. et al. (1999)
Unopposed production of granulocyte-macrophage colony-stimulating factor by tumors inhibits CD8
⁺
T cell responses by dysregulating antigen-presenting cell maturation.
J. Immunol. 162: 5728-5737
Movahedi, K. et al. (2008) Identification of discrete tumor-induced myeloid-derived suppressor cell subpopulations with distinct T cell-suppressive activity. Blood 111: 4233-4244

Data and images