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Figure 2 | Arthritis Research & Therapy

Figure 2

From: Flow cytometric characterization of freshly isolated and culture expanded human synovial cell populations in patients with chronic arthritis

Figure 2

Analysis of total synovium-derived cell population with four-color flow cytometry: representative sample. Cells are stained with 7-aminoactinomycin (7-AAD), CD45- fluoro-isothiocyanate (FITC), one phycoerythrin (PE)-conjugated antibody and one allophycocyanin (APC)-conjugated antibody or their isotypes. (a) Dye exclusion of debris and dead cells in forward scatter (FSC)/FL3 in contour plot (top) and dot plot (bottom). G1 gates for live cells. (b) Subgating of CD45-negative stromal (G2) and CD45-positive (G3) hematopoietic cell populations based on isotype staining with IgG1-FITC and cell granularity (side scatter). (c) Quantification of CD34-expression and CD90-expression within the stromal fraction. The gating for CD90-positive cells was based on IgG1-PE specifically measured on CD45-negative cells. (d) Quantification of CD14, CD3, and CD20-expression within G3. The gating for CD20-positive cells was performed on IgG1-PE measured on total cells. (e) Plots showing CD105-APC, CD73-PE, CD146-PE, and human leucocyte antigen (HLA)-DR-PE staining in the stromal fraction with their appropriate isotype controls.

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