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The pellet was subsequently washed with 10 ml of G-solution (500 ml HBSS (Life Technologies, BE10-508F) with 10% BSA (Sigma-Aldrich, 126615-25 ml) and 1% penicillinstreptomycin (Life Technologies, 15140122)) and resuspended in 5.5 ml of gradient solution comprising 15% Optiprep (5 ml 10% RPMI (Life Technologies, 11875093) + 3 ml of 40% Optiprep that was diluted from 60% Optiprep with G-solution (Sigma-Aldrich, D1556)) per sample, and placed on top of 2.5 ml of the gradient solution