Polymorphonuclear leukocyte isolation from venous blood of the dog

Year 2019, Volume: 3 Issue: 3, 75 - 78, 31.12.2019

Güneş Karakurt Kader Yıldız

https://doi.org/10.30704/http-www-jivs-net.621263

Abstract

The current study aimed to isolate neutrophils 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 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 a light microscope. 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 a fast technic for isolation of the neutrophils from venous blood samples of dogs.

 

Keywords

dog; venous blood; polymorph nuclear leucocyte; neutrophil; isolation

Supporting Institution

Scientific Research Coordination Unit of Kırıkkale University

Project Number

Project code: 2017 / 080

Thanks

This study was financially supported by Scientific Research Coordination Unit of Kırıkkale University (Project code: 2017 / 080).

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