{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T06:29:46Z","timestamp":1790922586381,"version":"4.1.0"},"reference-count":34,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T00:00:00Z","timestamp":1742256000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>Gastrointestinal inflammation leads to maldigestion and systemic diseases in poultry. To tackle the problem of the industry and to search for therapeutic candidates <jats:italic>in vitro<\/jats:italic> models are inevitable. Both immersion and air-liquid interface explant models are available, although there is limited information on the size-dependent applicability and response to different pathogen-associated molecular patterns (PAMPs) in the case of these model systems. The study aimed to compare the morphology and viability of miniature chicken gut explant cultures obtained with a biopsy punch to examine the size-dependent change over time. To verify the applicability of the model, pathogen-associated molecular patterns (PAMPs): flagellin, lipoteichoic acid (LTA) and polyinosinic polycytidylic acid (poly I:C) were applied to induce inflammation. The 2\u202fmm diameter explants showed a decrease in metabolic activity measured by CCK-8 assay after 12\u202fh and a significantly higher extracellular lactate dehydrogenase activity indicating cellular damage compared to the 1\u202fmm explants, supported by histological differences after 24\u202fh of culturing. After 12\u202fh of incubation, the 1.5\u202fmm explants retained columnar epithelial lining with moderate damage of the lamina propria (H&amp;amp;E and pan-cytokeratin staining). Exposure to 100\u202f\u03bcg\/mL poly I:C reduced the metabolic activity of the 1.5\u202fmm explants. LTA and poly I:C increased IFN-<jats:italic>\u03b3<\/jats:italic> concentration at both applied doses and IFN-<jats:italic>\u03b1<\/jats:italic> concentration was elevated by 50\u202f\u03bcg\/mL poly I:C treatment. Flagellin administration raised IL-2, IL-6, and RANTES levels, while higher LTA and poly I:C concentrations increased the IFN-<jats:italic>\u03b3<\/jats:italic>\/IL-10 ratio. According to the observations, the viability and integrity of the explants decreases with their size. After 12\u202fh, the 1.5\u202fmm diameter miniature chicken ileal explant stimulated with PAMPs can be an appropriate model to mimic diseases involving tissue damage and inflammation.<\/jats:p>","DOI":"10.3389\/fvets.2025.1484333","type":"journal-article","created":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T17:21:06Z","timestamp":1742318466000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Miniature chicken ileal explant culture to investigate the inflammatory response induced by pathogen-associated molecular patterns"],"prefix":"10.3389","volume":"12","author":[{"given":"G\u00e1bor","family":"M\u00e1tis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Csilla","family":"Seb\u0151k","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D\u00e1vid G.","family":"Horv\u00e1th","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rege Anna","family":"M\u00e1rton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e1t\u00e9","family":"Mackei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00falia","family":"V\u00f6r\u00f6sh\u00e1zi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1gnes","family":"Kem\u00e9ny","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zsuzsanna","family":"Neogr\u00e1dy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ilona","family":"Varga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrik","family":"Tr\u00e1j","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,3,18]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"S7","DOI":"10.1177\/014860719902300503","article-title":"Gut as the largest immunologic tissue","volume":"23","author":"Takahashi","year":"1999","journal-title":"JPEN J Parenter Enteral Nutr"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1038\/nri1057","article-title":"Anatomical basis of tolerance and immunity to intestinal antigens","volume":"3","author":"Mowat","year":"2003","journal-title":"Nat Rev Immunol"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"1910","DOI":"10.3390\/cells10081910","article-title":"Two sides to every question: attempts to activate chicken innate immunity in 2D and 3D hepatic cell cultures","volume":"10","author":"Seb\u0151k","year":"2021","journal-title":"Cells"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"172","DOI":"10.3390\/vaccines10020172","article-title":"Role of physiology, immunity, microbiota, and infectious diseases in the gut health of poultry","volume":"10","author":"Wickramasuriya","year":"2022","journal-title":"Vaccine"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"559","DOI":"10.1097\/00001574-200311000-00008","article-title":"Immunologic regulation of toll-like receptors in gut epithelium","volume":"19","author":"Abreu","year":"2003","journal-title":"Curr Opin Gastroenterol"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"572","DOI":"10.1038\/s41586-023-05915-x","article-title":"Organization of the human intestine at single-cell resolution","volume":"619","author":"Hickey","year":"2023","journal-title":"Nature"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1007\/s10565-011-9187-5","article-title":"Explant culture of gastrointestinal tissue: a review of methods and applications","volume":"27","author":"Randall","year":"2011","journal-title":"Cell Biol Toxicol"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"895871","DOI":"10.3389\/fncel.2022.895871","article-title":"Enteric glial cells in immunological disorders of the gut","volume":"16","author":"Liu","year":"2022","journal-title":"Front Cell Neurosci"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.jim.2016.07.004","article-title":"The culture of gut explants: a model to study the mucosal response","volume":"438","author":"Russo","year":"2016","journal-title":"J Immunol Methods"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"3096","DOI":"10.3382\/ps\/pex160","article-title":"Development of a chicken ileal explant culture model for measurement of gut inflammation induced by lipopolysaccharide","volume":"96","author":"Zhang","year":"2017","journal-title":"Poult Sci"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"942","DOI":"10.1007\/s11626-016-0060-y","article-title":"Development and evaluation of a porcine in vitro colon organ culture technique","volume":"52","author":"Costa","year":"2016","journal-title":"In Vitro Cell Dev Biol Anim"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1007\/978-1-4939-3603-8_5","article-title":"Organotypical tissue cultures from fetal and neonatal murine Colon","volume":"1422","author":"Neckel","year":"2016","journal-title":"Methods Mol Biol"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"795","DOI":"10.1007\/s00418-008-0405-z","article-title":"Organotypical tissue cultures from adult murine colon as an in vitro model of intestinal mucosa","volume":"129","author":"Bareiss","year":"2008","journal-title":"Histochem Cell Biol"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"409","DOI":"10.3390\/ani10030409","article-title":"Effects of acute heat stress on a newly established chicken hepatocyte\u2014nonparenchymal cell co-culture model","volume":"10","author":"Mackei","year":"2020","journal-title":"Animals"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"55347","DOI":"10.3791\/55347-v","article-title":"The ex vivo Colon organ culture and its use in antimicrobial host defense studies","volume":"120","author":"Udden","year":"2017","journal-title":"J Vis Exp"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"7584","DOI":"10.3390\/ijms21207584","article-title":"Intestinal explant cultures from gilthead seabream (Sparus aurata, L.) allowed the determination of mucosal sensitivity to bacterial pathogens and the impact of a plant protein diet","volume":"21","author":"Pe\u00f1aranda","year":"2020","journal-title":"Int J Mol Sci"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1002\/alr.20110","article-title":"Human sinonasal explant system for testing cytotoxicity of intranasal agents","volume":"2","author":"Lim","year":"2012","journal-title":"Int Forum Allergy Rhinol"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"1580","DOI":"10.1016\/j.tiv.2009.07.015","article-title":"Development of a pig jejunal explant culture for studying the gastrointestinal toxicity of the mycotoxin deoxynivalenol: histopathological analysis","volume":"23","author":"Kolf-Clauw","year":"2009","journal-title":"Toxicol In Vitro"},{"key":"ref19","volume-title":"R: A language and environment for statistical computing","year":"2020"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1016\/j.imbio.2012.02.002","article-title":"The hypoxic environment reprograms the cytokine\/chemokine expression profile of human mature dendritic cells","volume":"218","author":"Blengio","year":"2013","journal-title":"Immunobiology"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"676782","DOI":"10.3389\/fphys.2021.676782","article-title":"Hypoxia and inflammation: insights from high-altitude physiology","volume":"12","author":"Pham","year":"2021","journal-title":"Front Physiol"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"e13249","DOI":"10.1111\/aji.13249","article-title":"Cytokine mRNA and protein expression by cell cultures of epithelial ovarian cancer\u2014methodological considerations on the choice of analytical method for cytokine analyses","volume":"84","author":"Israelsson","year":"2020","journal-title":"Am J Reprod Immunol"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"727","DOI":"10.1017\/S0043933910000693","article-title":"Chicken toll-like receptors and their role in immunity","volume":"66","author":"Kannaki","year":"2010","journal-title":"Worlds Poult Sci J"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"1132","DOI":"10.1172\/JCI32140","article-title":"Mucosal IL-10 and TGF-beta play crucial roles in preventing LPS-driven, IFN-gamma-mediated epithelial damage in human colon explants","volume":"118","author":"Jarry","year":"2008","journal-title":"J Clin Invest"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"20659","DOI":"10.1038\/s41598-021-00096-x","article-title":"Lactobacillus rhamnosus and Bifidobacterium longum alleviate colitis and cognitive impairment in mice by regulating IFN-\u03b3 to IL-10 and TNF-\u03b1 to IL-10 expression ratios","volume":"11","author":"Ma","year":"2021","journal-title":"Sci Rep"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"203","DOI":"10.3390\/nu10020203","article-title":"Toll-like receptors: regulators of the immune response in the human gut","volume":"10","author":"Hug","year":"2018","journal-title":"Nutrients"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.jim.2018.08.009","article-title":"Immunological responsiveness of intestinal tissue explants and mucosal mononuclear cells to ex vivo stimulation","volume":"463","author":"Brand","year":"2018","journal-title":"J Immunol Methods"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"130","DOI":"10.1016\/j.phrs.2015.06.016","article-title":"Short-term effects of poly(I:C) on gut permeability","volume":"101","author":"Moyano-Porcile","year":"2015","journal-title":"Pharmacol Res"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"45270","DOI":"10.1038\/srep45270","article-title":"A primary human macrophage-enteroid co-culture model to investigate mucosal gut physiology and host-pathogen interactions","volume":"7","author":"Noel","year":"2017","journal-title":"Sci Rep"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"350","DOI":"10.1136\/gut.46.3.350","article-title":"Human intestinal epithelial cells secrete interleukin-1 receptor antagonist and interleukin-8 but not interleukin-1 or interleukin-6","volume":"46","author":"Daig","year":"2000","journal-title":"Gut"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"15.25.1","DOI":"10.1002\/0471142735.im1525s104","article-title":"Dextran sulfate sodium (DSS)-induced colitis in mice","volume":"104","author":"Chassaing","year":"2014","journal-title":"Curr Protoc Immunol"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"567","DOI":"10.1038\/mi.2016.14","article-title":"Gut mucosal DAMPs in IBD: from mechanisms to therapeutic implications","volume":"9","author":"Boyapati","year":"2016","journal-title":"Mucosal Immunol"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"2527","DOI":"10.1007\/s11259-024-10428-7","article-title":"Immunomodulatory properties of chicken cathelicidin-2 investigated on an ileal explant culture","volume":"48","author":"M\u00e1tis","year":"2024","journal-title":"Vet Res Commun"},{"key":"ref34","doi-asserted-by":"crossref","DOI":"10.2139\/ssrn.4685306","article-title":"Miniature chicken ileal explant culture to investigate the inflammatory response induced by pathogen associated molecular patterns","author":"M\u00e1tis","year":"2024"}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1484333\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,18]],"date-time":"2025-03-18T17:21:09Z","timestamp":1742318469000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1484333\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,18]]},"references-count":34,"alternative-id":["10.3389\/fvets.2025.1484333"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2025.1484333","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,18]]},"article-number":"1484333"}}