Ficoll-separated mononuclear cells from sepsis patients are contaminated with granulocytes

Intensive Care Medicine
Erica L T van den AkkerJan W Koper

Abstract

To determine the cell content and purity of Ficoll-separated peripheral blood mononuclear cells and granulocyte isolates in sepsis patients compared to healthy controls. Prospective study in the adult and pediatric intensive care departments of the Erasmus University Medical Center in the Netherlands. Three sepsis patients (two adults, one child) and four healthy controls. Blood leukocytes were separated by Ficoll into an interface and a bottom fraction. The cell content and purity was analyzed by cytospin and flow-cytometric immunofluorescence. In sepsis patients, the interface consisted of 11-52% mononuclear cells only, due to high contamination with granulocytes (48-89%). This was in contrast to a high proportion of mononuclear cells (88-100%) in healthy controls. The bottom fraction showed a cell purity of >or=92% polymorphonuclear granulocytes in sepsis patients as well as in healthy controls. Ficoll-separated leukocytes of sepsis patients are not suitable for studying mononuclear cells but can be used for studying granulocytes with high purity. The mononuclear cell fraction is highly contaminated with granulocytes. Additional separation techniques are necessary to obtain a pure cell fraction.

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Citations

Jan 1, 2011·Polish Journal of Veterinary Sciences·R Urban-ChmielA Wernicki
Apr 18, 2009·Intensive Care Medicine·Erica L T van den AkkerAnita C S Hokken-Koelega
Sep 29, 2009·The Journal of Heart and Lung Transplantation : the Official Publication of the International Society for Heart Transplantation·Johannes A GovaertJoseph C Wu
Aug 26, 2016·Immunological Reviews·Patrizia ScapiniMarco A Cassatella
Mar 15, 2020·Journal of Leukocyte Biology·Marwan HassaniNienke Vrisekoop
Feb 2, 2021·The Journal of Infectious Diseases·Nico A F JanssenFrank L van de Veerdonk

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