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Several Orthomyxoviridae and Coronavirinae respiratory viruses require cleavage of envelope proteins to acquire viral infectivity and consequently, need a host protease or the addition of exogenous trypsin for efficient propagation. Host TMPRSS2 is a key protease responsible for viral cleavage. Stable expression of human TMPRSS2 in African green monkey-derived Vero cells can enhance the porcine epidemic diarrhea virus. However, considering the narrow host tropism of viruses, a porcine cell line expressing pig TMPRSS2 could be optimal for replicating pig-derived viruses. Herein, we generated and evaluated a pig-derived PK-15 cell line stably expressing pig TMPRSS2. This cell line markedly (&gt;1000-fold) and specifically enhanced the growth of influenza viruses. Furthermore, we demonstrated the usefulness of a PK-15 cell line lacking the Stat2 gene with a stable expression of pig TMPRSS2 for efficient virus isolation from clinical samples in the presence of type I interferons. Therefore, PK-15 cells expressing pig TMPRSS2 could be a valuable and promising tool for virus isolation, vaccine production, and virological studies of TMPRSS2-dependent viruses.<\/jats:p>","DOI":"10.3390\/pathogens13010018","type":"journal-article","created":{"date-parts":[[2023,12,24]],"date-time":"2023-12-24T20:42:30Z","timestamp":1703450550000},"page":"18","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Generation of a Porcine Cell Line Stably Expressing Pig TMPRSS2 for Efficient Isolation of Swine Influenza Virus"],"prefix":"10.3390","volume":"13","author":[{"given":"Yuri L","family":"Tanaka","sequence":"first","affiliation":[{"name":"Laboratory of Veterinary Microbiology, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki 8892192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3577-9718","authenticated-orcid":false,"given":"Maya","family":"Shofa","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Microbiology, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki 8892192, Japan"},{"name":"Graduate School of Medicine and Veterinary Medicine, University of Miyazaki, Miyazaki 8891692, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Erika P","family":"Butlertanaka","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Microbiology, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki 8892192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4748-1545","authenticated-orcid":false,"given":"Ahmad Massoud","family":"Niazi","sequence":"additional","affiliation":[{"name":"Graduate School of Medicine and Veterinary Medicine, University of Miyazaki, Miyazaki 8891692, Japan"},{"name":"Laboratory of Veterinary Pathology, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki 8892192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takuya","family":"Hirai","sequence":"additional","affiliation":[{"name":"Graduate School of Medicine and Veterinary Medicine, University of Miyazaki, Miyazaki 8891692, Japan"},{"name":"Laboratory of Veterinary Pathology, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki 8892192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3063-0997","authenticated-orcid":false,"given":"Hirohisa","family":"Mekata","sequence":"additional","affiliation":[{"name":"Center for Animal Disease Control, University of Miyazaki, Miyazaki 8892192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2792-4399","authenticated-orcid":false,"given":"Akatsuki","family":"Saito","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Microbiology, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki 8892192, Japan"},{"name":"Graduate School of Medicine and Veterinary Medicine, University of Miyazaki, Miyazaki 8891692, Japan"},{"name":"Center for Animal Disease Control, University of Miyazaki, Miyazaki 8892192, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,12,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1186\/s13567-022-01091-4","article-title":"Comprehensive Evaluation of the Safety and Immunogenicity of a Gene-Deleted Variant Pseudorabies Virus Attenuated Vaccine","volume":"53","author":"Li","year":"2022","journal-title":"Vet. 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