Short communication, Note

https://doi.org/10.24099/vet.arhiv.160616

Combined dextran and ficoll separation yields pure populations of chicken peripheral blood mononuclear cells - short communication

Mladen Jergović ; Centre for Research and Transfer of Knowledge in Biotechnology, University of Zagreb, Zagreb, Croatia
Gordana Nedeljković ; Department of Poultry Diseases with Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia
Željko Cvetić ; Department of Poultry Diseases with Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia
Željko Gottstein ; Department of Poultry Diseases with Clinic, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia
Krešo Bendelja ; Centre for Research and Transfer of Knowledge in Biotechnology, University of Zagreb, Zagreb, Croatia

Abstract

The ficoll density gradient method, the most widely used method for isolating avian peripheral blood mononuclear cells (PBMCs), results in significant contamination with nucleated thrombocytes. This thrombocyte contamination can affect both immunophenotyping and functional assays of T cells. We used full blood samples from ten vaccinated male layer hybrid chickens for isolating PBMCs, and have developed a low cost, combined dextran and ficoll density gradient separation method which, in comparison to the standard ficoll density gradient separation, markedly reduces thrombocyte contamination of the isolated PBMCs. An ex vivo T cell proliferation assay showed an overall reduced background proliferation in unstimulated and stimulated samples after combined dextran and ficoll separation of PBMCs, resulting in a statistically significant difference upon in vitro NDV challenge

Keywords

dextran; ficoll; chicken; thrombocytes; peripheral blood mononuclear cells

Hrčak ID:

186223

URI

https://hrcak.srce.hr/186223

Publication date:

6.9.2017.

Article data in other languages: croatian

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