Polymorphonuclear leukocyte isolation from venous blood of the dogs

Güneş Karakurt [1] , Kader Yıldız [2]


Kırıkkale University, Graduate School of Health Science, Department of Parasitology Turkey


Karakurt G.: ORCID: 0000-0001-7564-516X ;  Yıldız K.: ORCID:0000-0001-5802-6156


 *Güneş Karakurt: gunes_karakurt@hotmail.com

Received: 17.09.2019,  Accepted: 13.12.2019,  Available online: 13.12.2019

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; polymorphonuclear leukocyte; neutrophil; isolation

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