Name: MDSC Detection Kit, human
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Product data and images for MDSC Detection Kit, human

Figures

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant

^®

Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45

⁺

cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD

^–

cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter

^high

(SSC

^high

) cells, which correspond to low density PMN-MDSCs (region R5), CD14

⁺

cells (region R6) and SSC

^low

CD14

^–

cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14

⁺

HLA-DR

^–

. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^–

CD33

^int

A) Gating strategy:
All events	Gated on R1
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
Gated on R2	Gated on R3
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
B) Identification of low density PMN cells (PMN-MDSCs):
Gated on R4	Gated on R5
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
C) Identification of M-MDSCs:
Gated on R6	Gated on R6
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
D) Identification of e-MDSCs:
Gated on R7	Gated on R7
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .

Specifications for MDSC Detection Kit, human

Overview

The MDSC Detection Kit is developed to allow standardization and simplification of flow cytometric analysis of the subpopulations of MDSCs described so far. For greater flexibility, the product configuration also allows the extension of the analysis panel with additional specificities of interest (e.g. CD10, CD66b, CD15, and LOX-1).

Detailed product information

Background information

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous group of mononuclear and polymorphonuclear myeloid cells with different immunosuppressive as well as disease promoting functions¹,². In healthy volunteers, they are found in very small numbers, but can expand considerably in pathological conditions. Depending on the type and stage of the disease, the MDSC population might be present in different maturation phases. In human, three major MDSC subsets can be identified: low density polymorphonuclear-MDSCs (PMN-MDSCs), also known as granulocytic MDSCs (G-MDSCs), monocytic- MDSCs (M-MDSCs), and early-MDSCs (e-MDSCs)³. Validated unambiguous phenotypic markers for MDSCs are still lacking and therefore various combinations of specificities are used for their identification⁴.

Applications

Detection and quantification of MDSC subsets in human PBMCs. This product might be used for analysis of other starting materials, such as whole blood, lysed whole blood, or single-cell suspensions from human tissues. For such applications, optional reagents might be required, such as Red Blood Cell Lysis Solution (10×) or additional fluorochrome-conjugated antibodies.

References for MDSC Detection Kit, human

Publications

Marigo, I. et al. (2010) Tumor-induced tolerance and immune suppression depend on the C/EBPβ transcription factor. Immunity 32(6): 790-802
Marini, O. et al. (2017)
Mature CD10
⁺
and immature CD10
⁻
neutrophils present in G-CSF–treated donors display opposite effects on T cells.
Blood 129(10): 1343-1356
Veglia, F. et al. (2018) Myeloid-derived suppressor cells coming of age. Nat. Immunol. 19(2): 108-119
Mandruzzato, S. et al. (2016) Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study. Cancer Immunol. Immunother. 65(2): 161-169
Gabrilovich, D. I. et al. (2012) Coordinated regulation of myeloid cells by tumours. Nat. Rev. Immunol. 12(4): 253-268
Bruger, A. M. et al. (2020) Protocol to assess the suppression of T-cell proliferation by human MDSC. Meth. Enzymol. 632: 155-192
Lang, S. et al. (2018) Clinical relevance and suppressive capacity of human myeloid-derived suppressor cell subsets. Clin Cancer Res. 24(19): 4834-4844
Bronte, V. et al. (2016) Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun. 7(1): 12150
Condamine, T. et al. (2016) Lectin-type oxidized LDL receptor-1 distinguishes population of human polymorphonuclear myeloid-derived suppressor cells in cancer patients. Sci Immunol 1(2): pii:aaf8943

Data and images

Data and images for MDSC Detection Kit, human

Figures

Figure 1

Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant

^®

Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45

⁺

^–

cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter

^high

(SSC

^high

) cells, which correspond to low density PMN-MDSCs (region R5), CD14

⁺

cells (region R6) and SSC

^low

CD14

^–

⁺

HLA-DR

^–

. D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^–

CD33

^int

A) Gating strategy:
All events	Gated on R1
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
Gated on R2	Gated on R3
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
B) Identification of low density PMN cells (PMN-MDSCs):
Gated on R4	Gated on R5
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
C) Identification of M-MDSCs:
Gated on R6	Gated on R6
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .
D) Identification of e-MDSCs:
Gated on R7	Gated on R7
View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .	View details Figure 1 Human PBMCs were stained using the MDSC Detection Kit, human and analyzed by flow cytometry using the MACSQuant ^® Analyzer 10. A) To exclude red blood cells and identify leukocytes, a first gate on CD45 ⁺ cells was set (region R1). Next, to eliminate doublets, a gate on single cells in Forward scatter-A (A=area) versus Forward scatter-H (H=height) was set (region R2). These cells were further distinguished from debris via Forward scatter-A and Side scatter-A (region R3). Afterwards, a gate on viable cells (7-AAD ^– cells) was set (region R4). B) Cells from region R4 were further separated into 3 subsets: side scatter ^high (SSC ^high ) cells, which correspond to low density PMN-MDSCs (region R5), CD14 ⁺ cells (region R6) and SSC ^low CD14 ^– cells (region R7). Low density PMN-MDSCs contained in region R5 were further separated into 4 subsets based on the expression of CD16 and CD11b (gates R8 to R11). C) Cells stained with MDSC Control Cocktail from R6 were displayed with CD14 versus REA Control-FITC, and a region enclosing >99.5% of cells was set (region R12). This region was transferred to the cells labeled with MDSC Staining Cocktail, depicting M-MDSCs as CD14 ⁺ HLA-DR ^– . D) Cells stained with MDSC Control Cocktail from region R7 were displayed with REA Control-FITC versus REA-Control-PE, and a region above the background staining, containing ^– CD33 ^int .

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Specifications for MDSC Detection Kit, human

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Publications

Marigo, I. et al. (2010) Tumor-induced tolerance and immune suppression depend on the C/EBPβ transcription factor. Immunity 32(6): 790-802
Marini, O. et al. (2017)
Mature CD10
⁺
and immature CD10
⁻
neutrophils present in G-CSF–treated donors display opposite effects on T cells.
Blood 129(10): 1343-1356
Veglia, F. et al. (2018) Myeloid-derived suppressor cells coming of age. Nat. Immunol. 19(2): 108-119
Mandruzzato, S. et al. (2016) Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study. Cancer Immunol. Immunother. 65(2): 161-169
Gabrilovich, D. I. et al. (2012) Coordinated regulation of myeloid cells by tumours. Nat. Rev. Immunol. 12(4): 253-268
Bruger, A. M. et al. (2020) Protocol to assess the suppression of T-cell proliferation by human MDSC. Meth. Enzymol. 632: 155-192
Lang, S. et al. (2018) Clinical relevance and suppressive capacity of human myeloid-derived suppressor cell subsets. Clin Cancer Res. 24(19): 4834-4844
Bronte, V. et al. (2016) Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun. 7(1): 12150
Condamine, T. et al. (2016) Lectin-type oxidized LDL receptor-1 distinguishes population of human polymorphonuclear myeloid-derived suppressor cells in cancer patients. Sci Immunol 1(2): pii:aaf8943

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MDSC Detection Kit, human

MDSC Detection Kit, human

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Specifications for MDSC Detection Kit, human

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Specifications

Specifications for MDSC Detection Kit, human

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