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Cell suspensions were passed through a 70 µm and 40 µm cell strainer (BD Bioscience), resuspended in 2.5 mL of 40% Percoll preparation (20 mL Percoll [GE Healthcare], 2.2 mL 10×PBS, 27.8 mL HBSS without Ca 2+ and Mg 2+ [Sigma-Aldrich]) and layered onto 1.5 mL 60% Percoll (30 mL Percoll, 3.3 mL 10×PBS, 16.7 mL HBSS)