{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T03:49:34Z","timestamp":1792813774801,"version":"4.5.0"},"reference-count":51,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,9,12]],"date-time":"2020-09-12T00:00:00Z","timestamp":1599868800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["240245660"],"award-info":[{"award-number":["240245660"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["415089553"],"award-info":[{"award-number":["415089553"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["278001972"],"award-info":[{"award-number":["278001972"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["272983813"],"award-info":[{"award-number":["272983813"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["416072091"],"award-info":[{"award-number":["416072091"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SFB 1029, project B01"],"award-info":[{"award-number":["SFB 1029, project B01"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Feline coronaviruses (FCoVs) infect both wild and domestic cat populations world-wide. FCoVs present as two main biotypes: the mild feline enteric coronavirus (FECV) and the fatal feline infectious peritonitis virus (FIPV). FIPV develops through mutations from FECV during a persistence infection. So far, the molecular mechanism of FECV-persistence and contributing factors for FIPV development may not be studied, since field FECV isolates do not grow in available cell culture models. In this work, we aimed at establishing feline ileum and colon organoids that allow the propagation of field FECVs. We have determined the best methods to isolate, culture and passage feline ileum and colon organoids. Importantly, we have demonstrated using GFP-expressing recombinant field FECV that colon organoids are able to support infection of FECV, which were unable to infect traditional feline cell culture models. These organoids in combination with recombinant FECVs can now open the door to unravel the molecular mechanisms by which FECV can persist in the gut for a longer period of time and how transition to FIPV is achieved.<\/jats:p>","DOI":"10.3390\/cells9092085","type":"journal-article","created":{"date-parts":[[2020,9,13]],"date-time":"2020-09-13T21:11:32Z","timestamp":1600031492000},"page":"2085","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Development of Feline Ileum- and Colon-Derived Organoids and Their Potential Use to Support Feline Coronavirus Infection"],"prefix":"10.3390","volume":"9","author":[{"given":"Gergely","family":"Tekes","sequence":"first","affiliation":[{"name":"Institute of Virology, Justus Liebig University Giessen, 35390 Giessen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosina","family":"Ehmann","sequence":"additional","affiliation":[{"name":"Bundeswehr Institute of Microbiology, 80937 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8614-4993","authenticated-orcid":false,"given":"Steeve","family":"Boulant","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany"},{"name":"Research Group \u201cCellular Polarity and Viral Infection\u201d, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5606-1297","authenticated-orcid":false,"given":"Megan L.","family":"Stanifer","sequence":"additional","affiliation":[{"name":"Department of Infectious Diseases, Molecular Virology, Heidelberg University Hospital, 69120 Heidelberg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"225","DOI":"10.1016\/j.jfms.2008.09.008","article-title":"A review of feline infectious peritonitis virus infection: 1963\u20132008","volume":"11","author":"Pedersen","year":"2009","journal-title":"J. Feline Med. Surg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2735","DOI":"10.1099\/vir.0.19129-0","article-title":"Persistence and transmission of natural type I feline coronavirus infection","volume":"84","author":"Addie","year":"2003","journal-title":"J. Gen. Virol."},{"key":"ref_3","first-page":"1449","article-title":"Serologic studies of naturally occurring feline infectious peritonitis","volume":"37","author":"Pedersen","year":"1976","journal-title":"Am. J. Vet. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1136\/vr.131.10.223-a","article-title":"Feline coronavirus antibodies in UK cats","volume":"131","author":"Sparkes","year":"1992","journal-title":"Vet. Rec."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1354\/vp.42-3-321","article-title":"Morphologic features and development of granulomatous vasculitis in feline infectious peritonitis","volume":"42","author":"Kipar","year":"2005","journal-title":"Vet. Pathol."},{"key":"ref_6","first-page":"2036","article-title":"Pathogenesis of feline infetious peritonitis: Pathologic changes and immunofluorescence","volume":"42","author":"Weiss","year":"1981","journal-title":"Am. J. Vet. Res."},{"key":"ref_7","first-page":"382","article-title":"Pathogenesis of feline infectious peritonitis: Nature and development of viremia","volume":"42","author":"Weiss","year":"1981","journal-title":"Am. J. Vet. Res."},{"key":"ref_8","first-page":"567","article-title":"Morphologic and physical characteristics of feline infectious peritonitis virus and its growth in autochthonous peritoneal cell cultures","volume":"37","author":"Pedersen","year":"1976","journal-title":"Am. J. Vet. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/bs.aivir.2016.08.002","article-title":"Feline Coronaviruses: Pathogenesis of Feline Infectious Peritonitis","volume":"96","author":"Tekes","year":"2016","journal-title":"Adv. Virus Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.vetimm.2014.09.001","article-title":"The influence of age and genetics on natural resistance to experimentally induced feline infectious peritonitis","volume":"162","author":"Pedersen","year":"2014","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.tvjl.2014.04.016","article-title":"An update on feline infectious peritonitis: Diagnostics and therapeutics","volume":"201","author":"Pedersen","year":"2014","journal-title":"Vet. J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.tvjl.2014.04.017","article-title":"An update on feline infectious peritonitis: Virology and immunopathogenesis","volume":"201","author":"Pedersen","year":"2014","journal-title":"Vet. J."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Kim, Y., Liu, H., Galasiti Kankanamalage, A.C., Weerasekara, S., Hua, D.H., Groutas, W.C., Chang, K.O., and Pedersen, N.C. (2016). Reversal of the Progression of Fatal Coronavirus Infection in Cats by a Broad-Spectrum Coronavirus Protease Inhibitor. PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1005531"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1587","DOI":"10.1111\/jvim.15780","article-title":"Antiviral treatment using the adenosine nucleoside analogue GS-441524 in cats with clinically diagnosed neurological feline infectious peritonitis","volume":"34","author":"Dickinson","year":"2020","journal-title":"J. Vet. Intern. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"108398","DOI":"10.1016\/j.vetmic.2019.108398","article-title":"Characterization of amino acid substitutions in feline coronavirus 3C-like protease from a cat with feline infectious peritonitis treated with a protease inhibitor","volume":"237","author":"Perera","year":"2019","journal-title":"Vet. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1177\/1098612X19825701","article-title":"Efficacy and safety of the nucleoside analog GS-441524 for treatment of cats with naturally occurring feline infectious peritonitis","volume":"21","author":"Pedersen","year":"2019","journal-title":"J. Feline Med. Surg."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.vetmic.2018.04.026","article-title":"The nucleoside analog GS-441524 strongly inhibits feline infectious peritonitis (FIP) virus in tissue culture and experimental cat infection studies","volume":"219","author":"Murphy","year":"2018","journal-title":"Vet. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1177\/1098612X17729626","article-title":"Efficacy of a 3C-like protease inhibitor in treating various forms of acquired feline infectious peritonitis","volume":"20","author":"Pedersen","year":"2018","journal-title":"J. Feline Med. Surg."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/0378-1135(91)90096-X","article-title":"Antigenic analysis of feline coronaviruses with monoclonal antibodies (MAbs): Preparation of MAbs which discriminate between FIPV strain 79-1146 and FECV strain 79-1683","volume":"28","author":"Hohdatsu","year":"1991","journal-title":"Vet. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/BF01310494","article-title":"Characterization of monoclonal antibodies against feline infectious peritonitis virus type II and antigenic relationship between feline, porcine, and canine coronaviruses","volume":"117","author":"Hohdatsu","year":"1991","journal-title":"Arch. Virol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1007\/978-1-4615-9373-7_36","article-title":"Pathogenic differences between various feline coronavirus isolates","volume":"173","author":"Pedersen","year":"1984","journal-title":"Adv. Exp. Med. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.vetmic.2008.06.007","article-title":"An update on canine coronaviruses: Viral evolution and pathobiology","volume":"132","author":"Decaro","year":"2008","journal-title":"Vet. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4508","DOI":"10.1128\/JVI.72.5.4508-4514.1998","article-title":"Feline coronavirus type II strains 79-1683 and 79-1146 originate from a double recombination between feline coronavirus type I and canine coronavirus","volume":"72","author":"Herrewegh","year":"1998","journal-title":"J. Virol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1007\/s11262-012-0864-0","article-title":"Full genome analysis of a novel type II feline coronavirus NTU156","volume":"46","author":"Lin","year":"2013","journal-title":"Virus Genes"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Terada, Y., Matsui, N., Noguchi, K., Kuwata, R., Shimoda, H., Soma, T., Mochizuki, M., and Maeda, K. (2014). Emergence of pathogenic coronaviruses in cats by homologous recombination between feline and canine coronaviruses. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0106534"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6994","DOI":"10.1128\/JVI.00023-12","article-title":"A reverse genetics approach to study feline infectious peritonitis","volume":"86","author":"Tekes","year":"2012","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1128\/JVI.02339-07","article-title":"Genome organization and reverse genetic analysis of a type I feline coronavirus","volume":"82","author":"Tekes","year":"2008","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1326","DOI":"10.1128\/JVI.01568-09","article-title":"Chimeric feline coronaviruses that encode type II spike protein on type I genetic background display accelerated viral growth and altered receptor usage","volume":"84","author":"Tekes","year":"2010","journal-title":"J. Virol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"396","DOI":"10.4161\/bioe.32133","article-title":"Tackling feline infectious peritonitis via reverse genetics","volume":"5","author":"Thiel","year":"2014","journal-title":"Bioengineered"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4528","DOI":"10.1128\/JVI.77.8.4528-4538.2003","article-title":"Switching species tropism: An effective way to manipulate the feline coronavirus genome","volume":"77","author":"Haijema","year":"2003","journal-title":"J. Virol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1387","DOI":"10.1126\/science.aaf5211","article-title":"Replication of human noroviruses in stem cell-derived human enteroids","volume":"353","author":"Ettayebi","year":"2016","journal-title":"Science"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"11255","DOI":"10.1128\/JVI.00947-10","article-title":"Culturing the unculturable: Human coronavirus HKU1 infects, replicates, and produces progeny virions in human ciliated airway epithelial cell cultures","volume":"84","author":"Pyrc","year":"2010","journal-title":"J. Virol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1190","DOI":"10.1126\/science.1234852","article-title":"Growing self-organizing mini-guts from a single intestinal stem cell: Mechanism and applications","volume":"340","author":"Sato","year":"2013","journal-title":"Science"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Ehmann, R., Kristen-Burmann, C., Bank-Wolf, B., Konig, M., Herden, C., Hain, T., Thiel, H.J., Ziebuhr, J., and Tekes, G. (2018). Reverse Genetics for Type I Feline Coronavirus Field Isolate to Study the Molecular Pathogenesis of Feline Infectious Peritonitis. MBio, 9.","DOI":"10.1128\/mBio.01422-18"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.virol.2019.08.030","article-title":"Generating and evaluating type I interferon receptor-deficient and feline TMPRSS2-expressing cells for propagating serotype I feline infectious peritonitis virus","volume":"537","author":"Mettelman","year":"2019","journal-title":"Virology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.virol.2018.08.022","article-title":"Characterizing replication kinetics and plaque production of type I feline infectious peritonitis virus in three feline cell lines","volume":"525","author":"Mettelman","year":"2018","journal-title":"Virology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1038\/s41564-019-0594-3","article-title":"Asymmetric distribution of TLR3 leads to a polarized immune response in human intestinal epithelial cells","volume":"5","author":"Stanifer","year":"2020","journal-title":"Nat. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1038\/nature07935","article-title":"Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche","volume":"459","author":"Sato","year":"2009","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1762","DOI":"10.1053\/j.gastro.2011.07.050","article-title":"Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett\u2019s epithelium","volume":"141","author":"Sato","year":"2011","journal-title":"Gastroenterology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"459","DOI":"10.3389\/fimmu.2017.00459","article-title":"Type I and Type III Interferons Display Different Dependency on Mitogen-Activated Protein Kinases to Mount an Antiviral State in the Human Gut","volume":"8","author":"Pervolaraki","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Triana, S., Stanifer, M.L., Shahraz, M., Mukenhirn, M., Kee, C., Ordo\u00f1ez-Rueda, D., Paulsen, M., Benes, V., Boulant, S., and Alexandrov, T. (2020, August 19). Single-cell transcriptomics reveals immune response of intestinal cell types to viral infection. bioRxiv Preprint Serv. Biol., Available online: https:\/\/doi.org\/10.1101\/2020.08.19.255893.","DOI":"10.1101\/2020.08.19.255893"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1016\/j.stem.2018.11.016","article-title":"Human Intestinal Organoids Maintain Self-Renewal Capacity and Cellular Diversity in Niche-Inspired Culture Condition","volume":"23","author":"Fujii","year":"2018","journal-title":"Cell Stem Cell"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/nmeth.1802","article-title":"Controlled gene expression in primary Lgr5 organoid cultures","volume":"9","author":"Koo","year":"2011","journal-title":"Nat. Methods"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1038\/nm.2470","article-title":"Isolation and in vitro expansion of human colonic stem cells","volume":"17","author":"Jung","year":"2011","journal-title":"Nat. Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"107863","DOI":"10.1016\/j.celrep.2020.107863","article-title":"Critical Role of Type III Interferon in Controlling SARS-CoV-2 Infection in Human Intestinal Epithelial Cells","volume":"32","author":"Stanifer","year":"2020","journal-title":"Cell Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1007\/s00441-018-2924-9","article-title":"Developing a 3D intestinal epithelium model for livestock species","volume":"375","author":"Derricott","year":"2019","journal-title":"Cell Tissue Res."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Kramer, N., Pratscher, B., Meneses, A.M.C., Tschulenk, W., Walter, I., Swoboda, A., Kruitwagen, H.S., Schneeberger, K., Penning, L.C., and Spee, B. (2020). Generation of Differentiating and Long-Living Intestinal Organoids Reflecting the Cellular Diversity of Canine Intestine. Cells, 9.","DOI":"10.3390\/cells9040822"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Chandra, L., Borcherding, D.C., Kingsbury, D., Atherly, T., Ambrosini, Y.M., Bourgois-Mochel, A., Yuan, W., Kimber, M., Qi, Y., and Wang, Q. (2019). Derivation of adult canine intestinal organoids for translational research in gastroenterology. BMC Biol., 17.","DOI":"10.1186\/s12915-019-0652-6"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/1297-9716-44-71","article-title":"Establishment of feline intestinal epithelial cell cultures for the propagation and study of feline enteric coronaviruses","volume":"44","author":"Desmarets","year":"2013","journal-title":"Vet. Res."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1698","DOI":"10.1099\/vir.0.020214-0","article-title":"Sites of feline coronavirus persistence in healthy cats","volume":"91","author":"Kipar","year":"2010","journal-title":"J. Gen. Virol."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Pervolaraki, K., Rastgou Talemi, S., Albrecht, D., Bormann, F., Bamford, C., Mendoza, J.L., Garcia, K.C., McLauchlan, J., Hofer, T., and Stanifer, M.L. (2018). Differential induction of interferon stimulated genes between type I and type III interferons is independent of interferon receptor abundance. PLoS Pathog., 14.","DOI":"10.1101\/448357"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/9\/2085\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:09:21Z","timestamp":1760177361000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/9\/2085"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,12]]},"references-count":51,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["cells9092085"],"URL":"https:\/\/doi.org\/10.3390\/cells9092085","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,12]]}}}