Combined dextran and ficoll separation yields pure populations of chicken peripheral blood mononuclear cells - short communicationia
| Autor: | Željko Cvetić, Gordana Nedeljković, Mladen Jergović, Krešo Bendelja, Željko Gottstein |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
General Veterinary Density gradient Chemistry T cell 0402 animal and dairy science Ficoll 04 agricultural and veterinary sciences 040201 dairy & animal science Peripheral blood mononuclear cell Molecular biology dextran ficoll chicken thrombocytes peripheral blood mononuclear cells 03 medical and health sciences chemistry.chemical_compound dextran ficoll chicken thrombocytes peripheral blood mononuclear cells 030104 developmental biology medicine.anatomical_structure Dextran Immunophenotyping medicine dekstran fikol pilići trombociti leukociti Platelet Ex vivo |
| Zdroj: | Veterinarski arhiv Volume 87 Issue 5 |
| ISSN: | 1331-8055 0372-5480 |
| Popis: | 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 Metoda razdvajanja gradijentom gustoće pomoću fikola najčešće je korištena metoda za izdvajanje leukocita iz periferne krvi ptica, ali koja rezultira značajnom kontaminacijom trombocitima. Trombociti u populaciji izdvojenih stanica utječu na rezultate fenotipskih (imunofenotipizacija), ali i funkcionalnih testova T limfocita. Koristeći uzorke pune krvi od 10 cijepljenih muških pilića komercijalnog hibrida nesilica razvili smo metodu izdvajanja leukocita kombiniranom primjenom dekstrana i gradijenta gustoće pomoću fikola CD8αex vivo test proliferacije T limfocita pokazao je smanjenu pozadinsku proliferaciju nestimuliranih stanica nakon izdvajanja leukocita kombiniranom primjenom dekstrana i fikola. |
| Databáze: | OpenAIRE |
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