Determination of poultry heterophile functions by flow cytometry

Year 2020, Vetexpo-2019, KHVD-2019, 32 - 32, 31.03.2020

Erdal Matur Mert Erek Ezgi Ergen Bilge Acar Bolat Mukaddes Özcan

Abstract

The aim of this study is to modify the flow cytometric methods for avian heterophil that used to analyze
neutrophil functions. Within the aim of the project, we tested the amount of blood for acquiring the
heterophil and the storage duration of the blood before the analysis and the time of centrifugation. Also,
we tested the amounts of cell suspension and dihydrordamine-123 (DHR-123) during the flow cytometric
analysis. We reviewed the amount of porbol miristat asetat (PMA) used to stimulate the oxidative burst
and the amount of formyl methionyl-leucyl-phenylalanine (fMLP) used to stimulate chemotaxic activity.
Experiments on the incubation temperature and incubation duration were also performed. The results
showed that 0.5-3 ml of blood could be used to detect heterophil functions and it would be ideal to study in
fresh blood samples. However, it also showed that the stored blood can be used for a maximum of 8 hours
at +4 degrees. In order to isolate the cells, centrifugation of blood samples for 30 minutes would be
sufficient, and it would be appropriate to use 30μL from the cell suspension. DHR-123, which is used as a
chemical probe to measure heterophil functions, had to be used in 2μL, and when used excessively, it
affected the heterophil functions negatively. In addition, it was seen that using 2μL each of fMLP, which is
used as an oxidative burst stimulant, and PMA as a stimulant of chemotaxic activity was sufficient. It was
concluded that the incubation at 41 ° C for 5 minutes after stimulating the heterophil would also be
sufficient. As a result, it was thought that this study could be used to isolate heterophil and to analyze with
flow cytometry and to contribute further research and clinical studies in poultry.

Keywords

Heterophil, flow cytometry

References

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