Polymorphonuclear leukocyte isolation from venous blood of the dog

Year 2020, Vetexpo-2019, KHVD-2019, 52 - 52, 31.03.2020

Güneş Karakurt Kader Yıldız

Abstract

The aim of the current study was to neutrophil isolation from venous blood samples of healthy dogs.
Venous blood samples were obtained from Venae cephalica of clinically healthy dogs (n:5) into
heparinized tubes. The blood samples (2 mL) and Percoll dilutions (45%, 54%, 63%, and 72%) prepared
with Hanks Balanced Salt Solution (HBSS) were layered into sterile tubes. After centrifuge, the
polymorphonuclear leukocytes (PMN) were aspirated between 63% and 72% interfaces of the Percoll
dilutions into tubes. The samples of PMN observed under light microscopy. Viability was detected
microscopically after stained with trypan blue dye. Diff-Quick staining was used to detect neutrophil
purity of the isolated PMN. In the present study, the neutrophils ratio was calculated as 92% of the isolated
polymorphonuclear cells. The neutrophil viability was calculated as 98% of PMNs isolated from the
venous blood samples of healthy dogs. In the present study, the Percoll gradient centrifugation (72%, 63%,
54% and 45%) is fast technic for isolation of the neutrophils from venous blood samples of dogs.

Keywords

dog, venous blood

References

• .