Assessment of Minimal Residual Disease in Ewing Sarcoma
Figure 1
Sequential gating to identify Ewing sarcoma cells. Cultured A673 cells undergo sequential gating to identify Ewing sarcoma cells. Mononuclear cells are separated from blood or marrow by density gradient centrifugation, stained with monoclonal antibodies (CD99 PE, CD45 FITC, CD14 PerCP, CD34 APC), exposed to anti-PE magnetic microbeads to enrich CD99+ cells using MACS technology (Miltenyi Biotec, Cologne, Germany), and then analyzed by flow cytometry. Analysis is performed using sequential gating strategy (gates 1–5) to purify the CD99 bright positive CD45 negative tumor cells as shown in this example of A673 Ewing sarcoma culture cells.
