title: Overnight storage of blood in ACD tubes at 4{degrees}C increases NK cell fraction in peripheral blood mononuclear cells.
abstract: A considerable variabilility in the effects of sample handling on NK cytotoxicity has been observed. Using flow cytometry, NK cytotoxicity assays and lymphocyte subset analysis of Ficoll-Hypaque-separated peripheral blood mononuclear cells (PBMCs) isolated from whole blood stored under various conditions were performed. The NK cytotoxicity of samples in heparin tubes stored overnight at 4 and 22°C, as well as at 22°C in acid citrate dextrose (ACD) tubes, was lower than that of a fresh sample. However, the NK cytotoxicity of samples in an ACD tube stored at 4°C was similar to that of a fresh sample. Based on lymphocyte subset analysis, samples in an ACD tube stored at 4°C showed a lower percentage of CD3+ T cells and a higher percentage of CD16/56+ NK cells compared to samples stored under other conditions. The NK cytotoxicity of fresh samples and samples in ACD tubes stored in a Styrofoam cooler box did not differ significantly; however, the differences were inconsistent. Overnight storage of peripheral blood in ACD tubes at 4°C is optimum for retention of NK cytotoxicity, the level of which is similar to that of fresh blood. This may be associated with an increased NK-cell fraction in Ficoll-Hypaque-separated PBMCs after overnight storage.
authors.affiliation: Research Center for Cancer Immunotherapy, Chonnam National University Hwasun Hospital, 160 Ilsimri, Hwasun-eup, Hwasun-gun, Jeollanam-do, 519-809, Korea.
authors.forename: Da-Woon
authors.initials: DW
authors.lastname: Kim
authors.forename: Youn-Young
authors.initials: YY
authors.lastname: Jang
authors.forename: Myung-Geun
authors.initials: MG
authors.lastname: Shin
authors.forename: Jong-Hee
authors.initials: JH
authors.lastname: Shin
authors.forename: Soon-Pal
authors.initials: SP
authors.lastname: Suh
authors.forename: Dong-Wook
authors.initials: DW
authors.lastname: Ryang
authors.forename: Meesun
authors.initials: M
authors.lastname: Yoon
authors.forename: Je-Jung
authors.initials: JJ
authors.lastname: Lee
authors.forename: Sang-Ki
authors.initials: SK
authors.lastname: Kim
authors.forename: Duck
authors.initials: D
authors.lastname: Cho
mesh_terms: D001793:Blood Preservation / Q000379:methods*; D001800:Blood Specimen Collection / Q000379:methods*; D002478:Cells, Cultured; D019343:Citric Acid / Q000737:chemistry* / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D005434:Flow Cytometry; D005947:Glucose / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D016131:Lymphocyte Subsets / Q000166:cytology / Q000187:drug effects / Q000276:immunology
chemical_list: C002113:acid citrate dextrose; D019343:Citric Acid; D005947:Glucose
keywords: Anticoagulant; acid citrate dextrose tube; cytotoxicity; natural killer cells; temperature