{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T11:17:03Z","timestamp":1788261423754,"version":"build-2803163510"},"reference-count":35,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004156","name":"Mahidol University","doi-asserted-by":"publisher","award":["FF-168\/2568"],"award-info":[{"award-number":["FF-168\/2568"]}],"id":[{"id":"10.13039\/501100004156","id-type":"DOI","asserted-by":"publisher"}]},{"award":["FF-168\/2568"],"award-info":[{"award-number":["FF-168\/2568"]}],"id":[{"id":"https:\/\/ror.org\/01znkr924","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Porcine epidemic diarrhea virus (PEDV) continues to cause devastating economic losses in the swine industry. However, the lack of species-specific in vitro models and reliance on costly animal models hinders development of effective interventions. We established a high-fidelity platform using porcine induced pluripotent stem cells (piPSCs; line VSMUi001-C) via a 28-day stepwise differentiation protocol. Successful lineage commitment was validated by OCT4 downregulation and robust FOXA2\/SOX17 expression at day 5. By day 28, the piPSC-derived intestinal epithelial cells (piPSC-IECs) exhibited organized LGR5+ niches, MUC2+ goblet cells, and Villin+ enterocytes, mirroring native tissue architecture. Barrier functionality was confirmed by the continuous distribution of the tight junction protein zonula occludens-1 (ZO-1). Our results demonstrate that piPSC-IECs are permissive to PEDV and support productive infection, encompassing the complete viral life cycle from entry to release of infectious progeny. Productive replication was confirmed by direct immunodetection of viral antigen within inoculated cells. Viral challenge induced characteristic cytopathic effects and a specific pro-inflammatory elevation of tumor necrosis factor-alpha (TNF-\u03b1). Notably, infection fragmented ZO-1, providing a mechanistic explanation for the leaky gut phenotype observed in vivo. This platform serves as a robust system for investigating viral kinetics and host\u2013pathogen interactions with high species-specific relevance.<\/jats:p>","DOI":"10.3390\/ani16172718","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T10:32:18Z","timestamp":1788258738000},"page":"2718","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Porcine iPSC-Derived Intestinal Cells as a High-Fidelity Platform for PEDV Pathogenesis"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4132-0142","authenticated-orcid":false,"given":"Tharathip","family":"Muangthong","sequence":"first","affiliation":[{"name":"Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"},{"name":"Pathobiology Information and Learning Center, Department of Pathobiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-8502-8615","authenticated-orcid":false,"given":"Warunya","family":"Chakritbudsabong","sequence":"additional","affiliation":[{"name":"Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"},{"name":"Laboratory of Cellular Biomedicine and Veterinary Medicine, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"given":"Jarupa","family":"Taowan","sequence":"additional","affiliation":[{"name":"The Monitoring and Surveillance Center for Zoonotic Diseases in Wildlife and Exotic Animals, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Phakhin","family":"Jantahiran","sequence":"additional","affiliation":[{"name":"Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"},{"name":"Laboratory of Cellular Biomedicine and Veterinary Medicine, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Wanatchaporn","family":"Jaroensuk","sequence":"additional","affiliation":[{"name":"Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"},{"name":"Laboratory of Cellular Biomedicine and Veterinary Medicine, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Kampon","family":"Kaeoket","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences and Public Health, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"}]},{"given":"Sasitorn","family":"Rungarunlert","sequence":"additional","affiliation":[{"name":"Department of Preclinic and Applied Animal Science, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"},{"name":"Laboratory of Cellular Biomedicine and Veterinary Medicine, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ibrahim, Y.M., Liu, C., Yu, Y., Yang, L., Chen, Q., Ma, W., Werid, G.M., Li, S., Luo, J., and Gao, S. (2026). Swine Enteric Coronaviruses: An Updated Overview of Epidemiology, Diagnosis, Prevention, and Control. Animals, 16.","DOI":"10.3390\/ani16030458"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.tvjl.2015.02.017","article-title":"Porcine epidemic diarrhea virus infection: Etiology, epidemiology, pathogenesis and immunoprophylaxis","volume":"204","author":"Jung","year":"2015","journal-title":"Vet. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"198045","DOI":"10.1016\/j.virusres.2020.198045","article-title":"Porcine epidemic diarrhea virus (PEDV): An update on etiology, transmission, pathogenesis, and prevention and control","volume":"286","author":"Jung","year":"2020","journal-title":"Virus Res."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Chen, Y., Zhou, J., Wang, X., Ma, L., Li, J., Yang, L., Yuan, H., Pang, D., and Ouyang, H. (2022). Porcine Epidemic Diarrhea Virus: An Updated Overview of Virus Epidemiology, Virulence Variation Patterns and Virus\u2013Host Interactions. Viruses, 14.","DOI":"10.3390\/v14112434"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e01682-18","DOI":"10.1128\/JVI.01682-18","article-title":"Porcine Intestinal Enteroids: A New Model for Studying Enteric Coronavirus Porcine Epidemic Diarrhea Virus Infection and the Host Innate Response","volume":"93","author":"Li","year":"2019","journal-title":"J. Virol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"104378","DOI":"10.1016\/j.micpath.2020.104378","article-title":"Porcine epidemic diarrhea virus infection blocks cell cycle and induces apoptosis in pig intestinal epithelial cells","volume":"147","author":"Shen","year":"2020","journal-title":"Microb. Pathog."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1038\/s41541-025-01157-2","article-title":"Characterization of a suspension Vero cell line for viral vaccine production","volume":"10","author":"Bourigault","year":"2025","journal-title":"npj Vaccines"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Gerber, P.F., Cao, D., Xiao, C.T., Chen, Q., Lager, K., Bosch, B.J., Meng, X.J., Halbur, P.G., and Opriessnig, T. (2023). Failure to experimentally infect 10 days-old piglets with a cell culture-propagated infectious stock of a classical genotype 1a porcine epidemic diarrhea virus. Front. Vet. Sci., 10.","DOI":"10.3389\/fvets.2023.1279162"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Ma, P., Fang, P., Ren, T., Fang, L., and Xiao, S. (2022). Porcine Intestinal Organoids: Overview of the State of the Art. Viruses, 14.","DOI":"10.3390\/v14051110"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Chakritbudsabong, W., Chaiwattanarungruengpaisan, S., Sariya, L., Pamonsupornvichit, S., Ferreira, J.N., Sukho, P., Gronsang, D., Tharasanit, T., Dinnyes, A., and Rungarunlert, S. (2021). Exogenous LIN28 Is Required for the Maintenance of Self-Renewal and Pluripotency in Presumptive Porcine-Induced Pluripotent Stem Cells. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.709286"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Neira, J.A., Conrad, J.V., Rusteika, M., and Chu, L.F. (2024). The progress of induced pluripotent stem cells derived from pigs: A mini review of recent advances. Front. Cell Dev. Biol., 12.","DOI":"10.3389\/fcell.2024.1371240"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1089\/cell.2012.0082","article-title":"Slow turning lateral vessel bioreactor improves embryoid body formation and cardiogenic differentiation of mouse embryonic stem cells","volume":"15","author":"Rungarunlert","year":"2013","journal-title":"Cell. Reprogram."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"17297","DOI":"10.1038\/srep17297","article-title":"A cost-effective system for differentiation of intestinal epithelium from human induced pluripotent stem cells","volume":"5","author":"Ogaki","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5989","DOI":"10.1038\/s41598-020-63151-z","article-title":"Generation of intestinal organoids derived from human pluripotent stem cells for drug testing","volume":"10","author":"Yoshida","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1016\/j.vetmic.2014.08.012","article-title":"Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene","volume":"173","author":"Oka","year":"2014","journal-title":"Vet. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e00202-17","DOI":"10.1128\/JVI.00202-17","article-title":"Tight Junction Protein Occludin Is a Porcine Epidemic Diarrhea Virus Entry Factor","volume":"91","author":"Luo","year":"2017","journal-title":"J. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"110124","DOI":"10.1016\/j.vetmic.2024.110124","article-title":"Nsp10-interacting host protein SAP18 restricts PEDV replication in Marc-145 cells via enhancing dephosphorylation of RIG-I","volume":"294","author":"Yang","year":"2024","journal-title":"Vet. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Zhang, M., Lv, L., Cai, H., Li, Y., Gao, F., Yu, L., Jiang, Y., Tong, W., Li, L., and Li, G. (2022). Long-Term Expansion of Porcine Intestinal Organoids Serves as an in vitro Model for Swine Enteric Coronavirus Infection. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.865336"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1038\/nature09691","article-title":"Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro","volume":"470","author":"Spence","year":"2011","journal-title":"Nature"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e23481","DOI":"10.1096\/fj.202302402R","article-title":"Induction of lung progenitor cell-like organoids by porcine pluripotent stem cells","volume":"38","author":"Yang","year":"2024","journal-title":"FASEB J."},{"key":"ref_21","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_22","doi-asserted-by":"crossref","unstructured":"Kang, Y., Park, H., Choe, B.H., and Kang, B. (2022). The Role and Function of Mucins and Its Relationship to Inflammatory Bowel Disease. Front. Med., 9.","DOI":"10.3389\/fmed.2022.848344"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2128","DOI":"10.1016\/j.febslet.2008.02.040","article-title":"Regulation of cell structure and function by actin-binding proteins: Villin\u2019s perspective","volume":"582","author":"Khurana","year":"2008","journal-title":"FEBS Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.virusres.2016.05.031","article-title":"Cellular entry of the porcine epidemic diarrhea virus","volume":"226","author":"Li","year":"2016","journal-title":"Virus Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1186\/s12985-018-1078-4","article-title":"Porcine epidemic diarrhea virus S1 protein is the critical inducer of apoptosis","volume":"15","author":"Chen","year":"2018","journal-title":"Virol. J."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1186\/s12985-024-02396-w","article-title":"Virological evaluation of natural and modified attapulgite against porcine epidemic diarrhoea virus","volume":"21","author":"Wang","year":"2024","journal-title":"Virol. J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1016\/j.vetmic.2015.03.022","article-title":"Structural alteration of tight and adherens junctions in villous and crypt epithelium of the small and large intestine of conventional nursing piglets infected with porcine epidemic diarrhea virus","volume":"177","author":"Jung","year":"2015","journal-title":"Vet. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wang, S., Wu, J., Wang, F., Wang, H., Wu, Z., Wu, S., and Bao, W. (2020). Expression Pattern Analysis of Antiviral Genes and Inflammatory Cytokines in PEDV-Infected Porcine Intestinal Epithelial Cells. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.00075"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Yuan, C., Sun, L., Chen, L., Li, L., Yao, Z., Wang, Y., Guo, H., Li, T., and Song, Q. (2022). Differences in cytokines expression between Vero cells and IPEC-J2 cells infected with porcine epidemic diarrhea virus. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.1002349"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Wu, Y., Wang, Y., Wang, X., Li, M., Yan, H., Shi, H., Shi, D., Chen, J., Guo, L., and Feng, L. (2024). Elevation of IL-8 secretion induced by PEDV infection via NF-\u03baB signaling pathway. Front. Cell. Infect. Microbiol., 14.","DOI":"10.3389\/fcimb.2024.1422560"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Bao, D., Yi, S., Zhao, L., Zhao, H., Liu, J., Wei, Y., Hu, G., and Liu, X. (2024). Porcine Epidemic Diarrhea Virus Infection of Porcine Intestinal Epithelial Cells Causes Mitochondrial DNA Release and the Activation of the NLRP3 Inflammasome to Mediate Interleukin-1\u03b2 Secretion. Vet. Sci., 11.","DOI":"10.3390\/vetsci11120643"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"109880","DOI":"10.1016\/j.virol.2023.109880","article-title":"PEDV promotes the differentiation of CD4+ T cells towards Th1, Tfh, and Treg cells via CD103+ DCs","volume":"587","author":"Yu","year":"2023","journal-title":"Virology"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Gabriel, V., Zdyrski, C., Sahoo, D.K., Ralston, A., Wickham, H., Bourgois-Mochel, A., Ahmed, B., Merodio, M.M., Paukner, K., and Pi\u00f1eyro, P. (2024). Adult Animal Stem Cell-Derived Organoids in Biomedical Research and the One Health Paradigm. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25020701"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2300543","DOI":"10.1002\/biot.202300543","article-title":"Cryopreservation of organoids: Strategies, innovation, and future prospects","volume":"19","author":"Han","year":"2024","journal-title":"Biotechnol. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1177\/2211068214561025","article-title":"TEER measurement techniques for in vitro barrier model systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J. Lab. Autom."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/17\/2718\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T10:37:40Z","timestamp":1788259060000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/17\/2718"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,1]]},"references-count":35,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ani16172718"],"URL":"https:\/\/doi.org\/10.3390\/ani16172718","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,1]]}}}