title: The effect of ex vivo refrigerated storage and cell preservation solution (Cyto-Chex II) on CD11b expression and oxidative burst activity of dog neutrophils.
abstract: The expression of CD11b and oxidative burst activity of dog neutrophils undergoing ex vivo refrigerated storage was studied using flow-cytometry . Additionally, the effect of a proprietary cell stabilization reagent (Cyto-Chex) on the expression of CD11b and oxidative burst activity was studied. Expression of CD11b was very high (>90% positive) on dog neutr ophils isolated from peripheral blood. Dog neutrophils showed a rapid and sustained increase in CD11b antigen density (P<0.01) during refrigerated storage, this increase was prevented by treatment with Cyto-Chex but was not completely blocked on the first day. There were no significant differences in mean antigen density between any days in the non-preserved group or between Days 1 to 4 in the Cyto-Chex treated group. The non-treated group showed significantly greater mean antigen density at all time points when compared to the preservative treated group (P<0.0001). Treatment with Cyto-Chex did not interfere with measurement of oxidative burst function on the first 2 days. Alterations of both resting oxidative activity and stimulated response were observed over time in both treated and untreated blood samples. Cyto-Chex treated samples showed a dramatic, significant decline in stimulated response after the third day of storage (P<0.001), while non-treated cells showed steadily increasing, but non-significant differences in stimulated response. Cyto-Chex was demonstrated to be a useful reagent for stabilization of dog neutrophil membrane antigens during storage, however this reagent is not recommended for preservation of cells for functional assays.
authors.affiliation: The Gastrointestinal Laboratory, Small Animal Medicine and Surgery, Texas A&M University, College Station, TX 77843-4474, USA. cruaux@cvm.tamu.edu
authors.forename: C G
authors.initials: CG
authors.lastname: Ruaux
authors.forename: D A
authors.initials: DA
authors.lastname: Williams
mesh_terms: D000818:Animals; D004285:Dogs / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D016177:Macrophage-1 Antigen / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D008297:Male; D009504:Neutrophils / Q000737:chemistry / Q000276:immunology*; D011309:Preservation, Biological / Q000379:methods / Q000662:veterinary*; D012034:Refrigeration; D016897:Respiratory Burst / Q000276:immunology; D013997:Time Factors
chemical_list: D016177:Macrophage-1 Antigen