{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:39:39Z","timestamp":1760236779178,"version":"build-2065373602"},"reference-count":51,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,12,19]],"date-time":"2021-12-19T00:00:00Z","timestamp":1639872000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Federal Ministry of Education and Research","doi-asserted-by":"publisher","award":["FKZ 031B0743"],"award-info":[{"award-number":["FKZ 031B0743"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Knowledge about the modes of action of immunomodulating compounds such as pathogens, drugs, or feed additives, e.g., probiotics, gained through controlled but animal-related in vitro systems using primary cultured peripheral blood mononuclear cells (PBMCs) will allow the development of targeted nutrition strategies. Moreover, it could contribute to the prevention of infectious diseases and the usage of antimicrobials, and further promote the health of the animals. However, to our knowledge, a protocol for the isolation of PBMCs with reduced thrombocyte count from chicken blood and subsequent cell culture over several days to assess the effects of immunomodulating compounds is not available. Therefore, we established an optimized protocol for blood sampling and immune cell isolation, culture, and phenotyping for chicken PBMCs. For blood sampling commercial Na\u2013citrate tubes revealed the highest count of vital cells compared to commercial Li\u2013heparin (p &lt; 0.01) and K3EDTA (p &lt; 0.05) tubes. Using combined dextran and ficoll density gradient separation, the thrombocyte count was significantly reduced (p &lt; 0.01) compared to slow-speed centrifugation with subsequent ficoll. For cell culture, the supplementation of RPMI-1640 medium with 10% chicken serum resulted in the lowest relative cell count of thrombocytes compared to fetal calf serum (FCS) (p &lt; 0.05). To validate the ability of the cell culture system to respond to stimuli, concanavalin A (conA) was used as a positive control. The optimized protocol allows the isolation and cultivation of vital PBMCs with reduced thrombocyte count from chicken blood for subsequent investigation of the modes of action of immunomodulating compounds.<\/jats:p>","DOI":"10.3390\/ani11123600","type":"journal-article","created":{"date-parts":[[2021,12,19]],"date-time":"2021-12-19T20:41:55Z","timestamp":1639946515000},"page":"3600","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Chicken Immune Cell Assay to Model Adaptive Immune Responses In Vitro"],"prefix":"10.3390","volume":"11","author":[{"given":"Filip","family":"Larsberg","sequence":"first","affiliation":[{"name":"Albrecht Daniel Thaer-Institute, Breeding Biology and Molecular Genetics, Humboldt University of Berlin, Unter den Linden 6, 10099 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maximilian","family":"Sprechert","sequence":"additional","affiliation":[{"name":"Albrecht Daniel Thaer-Institute, Breeding Biology and Molecular Genetics, Humboldt University of Berlin, Unter den Linden 6, 10099 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5915-8003","authenticated-orcid":false,"given":"Deike","family":"Hesse","sequence":"additional","affiliation":[{"name":"Albrecht Daniel Thaer-Institute, Breeding Biology and Molecular Genetics, Humboldt University of Berlin, Unter den Linden 6, 10099 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4387-2947","authenticated-orcid":false,"given":"Gudrun A.","family":"Brockmann","sequence":"additional","affiliation":[{"name":"Albrecht Daniel Thaer-Institute, Breeding Biology and Molecular Genetics, Humboldt University of Berlin, Unter den Linden 6, 10099 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9425-3356","authenticated-orcid":false,"given":"Susanne","family":"Kreuzer-Redmer","sequence":"additional","affiliation":[{"name":"Institute of Animal Nutrition and Functional Plant Compounds, University of Veterinary Medicine Vienna, Veterin\u00e4rplatz 1, 1210 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"26","DOI":"10.3389\/fvets.2019.00026","article-title":"Effects of Dietary Supplementation with Enterococcus faecium and Clostridium butyricum, Either Alone Or in Combination, on Growth and Fecal Microbiota Composition of Post-Weaning Pigs at a Commercial Farm","volume":"6","author":"Sato","year":"2019","journal-title":"Front. 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