title: Characterization of buffy coat-derived granulocytes for clinical use: a comparison with granulocyte colony-stimulating factor/dexamethasone-pretreated donor-derived products.
abstract: OBJECTIVE
  Buffy coat-derived granulocytes have been described as an alternative to the apheresis product from donors pretreated with dexamethasone and granulocyte colony-stimulating factor (G-CSF). The latter is - dependent on the local and national settings - obtained following a demanding and time-consuming procedure, which is undesirable in critically ill septic patients. In contrast, buffy coat-derived products have a large volume and are often heavily contaminated with red cells and platelets. We developed a new pooled buffy coat-derived product with high purity and small volume, and performed a comprehensive functional characterization of these granulocytes.
  
  
  METHODS
  We pooled ten buffy coats following the production of platelet concentrates. Saline 0·9% was added to decrease the viscosity and the product was split into plasma, red cells and a 'super' buffy coat. Functional data of the granulocytes were compared to those obtained with granulocytes from healthy controls and G-CSF/dexamethasone-pretreated donors.
  
  
  RESULTS
  Buffy coat-derived granulocytes showed adhesion, chemotaxis, reactive oxygen species production, degranulation, NETosis and in vitro killing of Staphylococcus aureus, Escherichia coli and Aspergillus species comparable to control and G-CSF/dexamethasone-derived granulocytes. Candida killing was superior compared to G-CSF/dexamethasone-derived granulocytes. Immunophenotyping was normal; especially no signs of activation in the buffy coat-derived granulocytes were seen. Viability was reduced. Buffy coats are readily available in the regular blood production process and would take away the concerns around the apheresis product.
  
  
  CONCLUSIONS
  The product described appears a promising alternative for transfusion purposes.
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: van de Geer
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.
authors.forename: R P
authors.initials: RP
authors.lastname: Gazendam
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.
authors.forename: A T J
authors.initials: AT
authors.lastname: Tool
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.
authors.forename: J L
authors.initials: JL
authors.lastname: van Hamme
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.|Department of Product and Process Development, Sanquin Blood Bank, Amsterdam, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: de Korte
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.
authors.forename: T K
authors.initials: TK
authors.lastname: van den Berg
authors.affiliation: Department of Immunopathology, Sanquin Research, Amsterdam, The Netherlands.|Department of Hematology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: S S
authors.initials: SS
authors.lastname: Zeerleder
authors.affiliation: Department of Blood Cell Research, Sanquin Research, Amsterdam, The Netherlands.|Department of Pediatric Hematology, Immunology & Infectious disease, Emma Children's Hospital, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: T W
authors.initials: TW
authors.lastname: Kuijpers
mesh_terms: D000328:Adult; D000954:Antigens, Surface / Q000378:metabolism; D058621:Blood Buffy Coat / Q000166:cytology*; D001781:Blood Component Removal; D001782:Blood Donors; D001792:Blood Platelets / Q000166:cytology; D002448:Cell Adhesion / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology*; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006098:Granulocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D007958:Leukocyte Count; D008297:Male; D019255:NADPH Oxidases / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000954:Antigens, Surface; D017382:Reactive Oxygen Species; D016179:Granulocyte Colony-Stimulating Factor; D003907:Dexamethasone; D019255:NADPH Oxidases
keywords: buffy coat; dexamethasone; granulocyte; granulocyte colony-stimulating factor; neutrophil; transfusion