TY - JOUR

T1 - Chemotaxis of canine polymorphonuclear neutrophil granulocytes using the under-agarose method applied to glass microscope slides

AU - Petersen, T. K.

AU - Jensen, A. L.

AU - Aaes, H.

PY - 1998

Y1 - 1998

N2 - The under-agarose chemotaxis assay method applied to glass microscope slides was optimised for canine polymorphonuclear granulocytes (PMN) with gelatin as protein supplement. Optimum chemotactic response occurred after approximately 135 min of incubation at 37°C with 5 x 105 cells/well and zymosan-activated serum (ZAS) as chemoattractant. Chemotactic (C) and random migration (R) were stated separately for each dog. Storage of blood samples for 90 min before isolation of PMN did not influence the C or R values (p > 0.05). The preanalytical variation was investigated and the proportion of variance, expressed as a percentage, due to each source for the C and R values, respectively, was determined: dog, 92%, 96%; blood sample, 0.4%, 0%; interslide, 3%, 2% and intraslide, 3%, 3%. The mean chemotactic responsiveness against ZAS and ZAS diluted 1:1 with RPMI 1640 (dZAS) was determined using 15 laboratory beagle dogs: C(ZAS) = 0.67 mm (range: 0.25-2.33 mm), coefficient of variation (CV) = 24%, C(dZAS) = 0.47 mm (range: 0.18-2.12 mm), CV = 22%. Also, the mean random migration was determined: 0.11 mm (range. 0.05-0.34 mm), CV = 25%. Neither age nor sex had any significant influence on chemotactic or random migration (p > 0.05). The effect of heat-inactivated pooled canine serum (HI-serum) incorporated into agarose was examined and it was found that this had a significant stimulatory effect on both chemotactic and random migratory responses (p < 0.05). Therefore, agarose with HI-serum is not useful when investigating the chemotactic function.

AB - The under-agarose chemotaxis assay method applied to glass microscope slides was optimised for canine polymorphonuclear granulocytes (PMN) with gelatin as protein supplement. Optimum chemotactic response occurred after approximately 135 min of incubation at 37°C with 5 x 105 cells/well and zymosan-activated serum (ZAS) as chemoattractant. Chemotactic (C) and random migration (R) were stated separately for each dog. Storage of blood samples for 90 min before isolation of PMN did not influence the C or R values (p > 0.05). The preanalytical variation was investigated and the proportion of variance, expressed as a percentage, due to each source for the C and R values, respectively, was determined: dog, 92%, 96%; blood sample, 0.4%, 0%; interslide, 3%, 2% and intraslide, 3%, 3%. The mean chemotactic responsiveness against ZAS and ZAS diluted 1:1 with RPMI 1640 (dZAS) was determined using 15 laboratory beagle dogs: C(ZAS) = 0.67 mm (range: 0.25-2.33 mm), coefficient of variation (CV) = 24%, C(dZAS) = 0.47 mm (range: 0.18-2.12 mm), CV = 22%. Also, the mean random migration was determined: 0.11 mm (range. 0.05-0.34 mm), CV = 25%. Neither age nor sex had any significant influence on chemotactic or random migration (p > 0.05). The effect of heat-inactivated pooled canine serum (HI-serum) incorporated into agarose was examined and it was found that this had a significant stimulatory effect on both chemotactic and random migratory responses (p < 0.05). Therefore, agarose with HI-serum is not useful when investigating the chemotactic function.

KW - Canine

KW - Chemotaxis

KW - Gelatin

KW - Neutrophil

KW - Under-agarose

UR - http://www.scopus.com/inward/record.url?scp=0031938674&partnerID=8YFLogxK

U2 - 10.1007/BF02628102

DO - 10.1007/BF02628102

M3 - Journal article

AN - SCOPUS:0031938674

VL - 8

SP - 31

EP - 36

JO - Comparative Haematology International

JF - Comparative Haematology International

SN - 0938-7714

IS - 1

ER -