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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.

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