Isolation of whole mononuclear cells from peripheral blood by the Ficoll Hypaque gradient

This simple and rapid method takes advantage of the density differences between mononuclear cells and other elements found in blood samples. Mononuclear cells and platelets collect on top of the Ficoll-Hypaque layer as they have a lower density. In contrast, red blood cells and granulocytes have a higher density than Ficoll-Hypaque and collect at the bottom of Ficoll-Hypaque layer. Platelets are separated from the mononuclear cells by subsequent washing or centrifugation through a FCS cushion gradient, which allows penetration of mononuclear cells but not platelets.

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