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.
Highlights
Polymorphonuclear leukocyte (PMN), granular leukocyte or granulocyte, is a type of white blood cell
There are many neutrophil isolation PMNs were detected between 63% and 72%
Methods, these methods do not apply to all animals interfaces of the Percoll dilutions after centrifuging
Summary
Polymorphonuclear leukocyte (PMN), granular leukocyte or granulocyte, is a type of white blood cell. The PMN consist of some granulocytic cells called neutrophils, eosinophils, and basophils. Most of the PMN are neutrophils, which ratio varies depending on the animal species. The neutrophils can live 3-12 hours in the organism, which can be as long as 1-2 days (Kruger et al, 2015). Cytokines and bacterial products prolong the life span of neutrophils. If it does not enter the process of inflammation, neutrophils are phagocytosed by macrophages in the organism (Mayadas et al, 2014). The innate immune system is a first defence mechanism that provides rapid protection after confronted with infectious microorganisms in the body (Kaplan and Radic, 2012). Neutrophils are one of the most important components of innate immunity.
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