{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,27]],"date-time":"2026-10-27T13:10:47Z","timestamp":1793106647675,"version":"4.5.0"},"reference-count":30,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,8,10]],"date-time":"2021-08-10T00:00:00Z","timestamp":1628553600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,8,10]],"date-time":"2021-08-10T00:00:00Z","timestamp":1628553600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Bovine-derived cultured cells, including Madin-Darby bovine kidney cells, are used worldwide; however, lipofection tend to result in low transfection efficiency, which has impeded the progress of veterinary research. We performed experiments to confirm the lipofection efficiency of bovine-derived cultured cells, to identify cells that suitable for lipofection. Several bovine tissues (endometrium, testis, ear tissue and foetal muscle) were collected, and primary cultured cells were prepared. Lipofection assay showed that only bovine endometrium (BE)-derived cells could be transfected efficiently (50\u201270%). BE cells can be divided into at least two types of cell populations (BE-1 and BE-2). The BE-1 cells, which were suitable for lipofection, were obtained by passages at short intervals and were negative for cytokeratin- and positive for vimentin-expression; the BE-2 cells did not have these characteristics and were not suitable for lipofection. Furthermore, the BE-1 cells and artificially immortalised cells of BE-1, iBE-1 cells, were utilised in a reporter assay requiring the introduction of multiple DNAs. Endometrial tissues can be collected from living cows, and BE-1 cells can be obtained easily by controlling passaging timing. The production of BE-1 cells and sharing the methods required to prepare them will contribute to the development of veterinary research.<\/jats:p>","DOI":"10.1038\/s41598-021-95848-0","type":"journal-article","created":{"date-parts":[[2021,8,10]],"date-time":"2021-08-10T10:03:10Z","timestamp":1628589790000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Bovine endometrium-derived cultured cells are suitable for lipofection"],"prefix":"10.1038","volume":"11","author":[{"given":"Mai","family":"Shiokawa","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryotaro","family":"Miura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aki","family":"Okubo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujiro","family":"Hagita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Itaru","family":"Yoshimura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Aoki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,8,10]]},"reference":[{"key":"95848_CR1","doi-asserted-by":"publisher","first-page":"3173","DOI":"10.1007\/s00216-010-3821-6","volume":"397","author":"TK Kim","year":"2010","unstructured":"Kim, T. K. & Eberwine, J. H. Mammalian cell transfection: The present and the future. Anal. Bioanal. Chem. 397, 3173\u20133178 (2010).","journal-title":"Anal. Bioanal. Chem."},{"key":"95848_CR2","doi-asserted-by":"publisher","first-page":"434","DOI":"10.1016\/0042-6822(65)90126-1","volume":"27","author":"A Vaheri","year":"1965","unstructured":"Vaheri, A. & Pagano, J. S. Infectious poliovirus RNA: A sensitive method of assay. Virology 27, 434\u2013436 (1965).","journal-title":"Virology"},{"key":"95848_CR3","first-page":"351","volume":"41","author":"JH McCutchan","year":"1968","unstructured":"McCutchan, J. H. & Pagano, J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J. Natl. Cancer Inst. 41, 351\u2013357 (1968).","journal-title":"J. Natl. Cancer Inst."},{"key":"95848_CR4","doi-asserted-by":"publisher","first-page":"456","DOI":"10.1016\/0042-6822(73)90341-3","volume":"52","author":"FL Graham","year":"1973","unstructured":"Graham, F. L. & van der Alex, J. E. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52, 456\u2013467 (1973).","journal-title":"Virology"},{"key":"95848_CR5","first-page":"2745","volume":"7","author":"C Chen","year":"1987","unstructured":"Chen, C. & Okayama, H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol. Cell Biol. 7, 2745\u20132752 (1987).","journal-title":"Mol. Cell Biol."},{"key":"95848_CR6","doi-asserted-by":"publisher","first-page":"7413","DOI":"10.1073\/pnas.84.21.7413","volume":"84","author":"PL Felgner","year":"1987","unstructured":"Felgner, P. L. et al. Lipofection: A highly efficient, lipid-mediated DNA-transfection procedure. Proc. Natl. Acad. Sci. 84, 7413\u20137417 (1987).","journal-title":"Proc. Natl. Acad. Sci."},{"key":"95848_CR7","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1186\/s12860-020-00254-5","volume":"21","author":"I Hyder","year":"2020","unstructured":"Hyder, I., Eghbalsaied, S. & Kues, W. A. Systematic optimization of square-wave electroporation conditions for bovine primary fibroblasts. BMC Mol. Cell. Biol. 21, 9 (2020).","journal-title":"BMC Mol. Cell. Biol."},{"key":"95848_CR8","doi-asserted-by":"publisher","first-page":"200","DOI":"10.1016\/j.gene.2017.05.025","volume":"30","author":"JS Osorio","year":"2017","unstructured":"Osorio, J. S. & Bionaz, M. Plasmid transfection in bovine cells: Optimization using a realtime monitoring of green fluorescent protein and effect on gene reporter assay. Gene 30, 200\u2013208 (2017).","journal-title":"Gene"},{"key":"95848_CR9","doi-asserted-by":"publisher","first-page":"574","DOI":"10.3181\/00379727-98-24111","volume":"98","author":"SH Mardin","year":"1958","unstructured":"Mardin, S. H. & Darby, N. B. Jr. Established kidney cell lines of normal adult bovine and ovine origin. Proc. Soc. Exp. Biol. Med. 98, 574\u2013576 (1958).","journal-title":"Proc. Soc. Exp. Biol. Med."},{"key":"95848_CR10","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1016\/0014-4827(91)90422-Q","volume":"197","author":"HT Huynh","year":"1991","unstructured":"Huynh, H. T., Robitaille, G. & Turner, J. D. Establishment of bovine mammary epithelial cells (MAC-T): An in vitro model for bovine lactation. Exp. Cell. Res. 197, 191\u2013199 (1991).","journal-title":"Exp. Cell. Res."},{"key":"95848_CR11","doi-asserted-by":"publisher","first-page":"34","DOI":"10.17533\/udea.rccp.v32n1a04","volume":"32","author":"AG Curcio","year":"2018","unstructured":"Curcio, A. G. et al. Efficiency of transgene expression in bovine cells varies according to cell type and gene transfer method. Rev. Colomb. Cienc. Pec. 32, 34\u201342 (2018).","journal-title":"Rev. Colomb. Cienc. Pec."},{"key":"95848_CR12","doi-asserted-by":"publisher","first-page":"177","DOI":"10.11620\/IJOB.2014.39.4.177","volume":"39","author":"L Jin","year":"2014","unstructured":"Jin, L., Kim, D. & Roh, S. Comparison of various transfection methods in human and bovine cultured cells. Int. J. Oral. Biol. 39, 177\u2013185 (2014).","journal-title":"Int. J. Oral. Biol."},{"key":"95848_CR13","first-page":"185","volume":"30","author":"RR Oliveira","year":"2005","unstructured":"Oliveira, R. R. et al. Effectiveness of liposomes to transfect livestock fibroblasts. Genet. Mol. Res. 30, 185\u2013196 (2005).","journal-title":"Genet. Mol. Res."},{"key":"95848_CR14","unstructured":"Bondzio, A., Walk, N., Sch\u00f6n, J. & Einspanier, R. Optimization of TransFectin Lipid Reagent-mediated transfection for different cell types. Bio-Rad Laboratories, Inc. tech note. 3197, (2005)."},{"key":"95848_CR15","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1016\/j.vascn.2004.08.014","volume":"51","author":"P Thomas","year":"2005","unstructured":"Thomas, P. & Smart, T. G. HEK293 cell line: A vehicle for the expression of recombinant proteins. J. Pharmacol. Tox. Met. 51, 187\u2013200 (2005).","journal-title":"J. Pharmacol. Tox. Met."},{"key":"95848_CR16","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.virol.2019.09.016","volume":"538","author":"M Shiokawa","year":"2019","unstructured":"Shiokawa, M. et al. END-phenomenon negative bovine viral diarrhea virus that induces the host\u2019s innate immune response supports propagation of BVDVs with different immunological properties. Virology 538, 97\u2013110 (2019).","journal-title":"Virology"},{"key":"95848_CR17","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1530\/jrf.0.0830239","volume":"83","author":"MA Fortier","year":"1988","unstructured":"Fortier, M. A., Guilbault, L. A. & Grasso, F. Specific properties of epithelial and stromal cells from the endometrium of cows. J. Reprod. Fertil. 83, 239\u2013248 (1988).","journal-title":"J. Reprod. Fertil."},{"key":"95848_CR18","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1007\/s11259-020-09770-3","volume":"44","author":"P Kelly","year":"2020","unstructured":"Kelly, P., Barry-Reidy, A., Brewer, A., Meade, K. G. & Farrelly, O. C. Improved filtration method to isolate pure populations of primary bovine endometrial epithelial and stromal cells for immunological studies. Vet. Res. Commun. 44, 29\u201339 (2020).","journal-title":"Vet. Res. Commun."},{"key":"95848_CR19","doi-asserted-by":"publisher","first-page":"531","DOI":"10.1262\/jrd.49.531","volume":"49","author":"S Murakami","year":"2003","unstructured":"Murakami, S., Shibaya, M., Takeuchi, K., Skarzynski, D. J. & Okuda, K. A passage and storage system for isolated bovine endometrial epithelial and stromal cells. J. Reprod. Dev. 49, 531\u2013538 (2003).","journal-title":"J. Reprod. Dev."},{"key":"95848_CR20","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.ymeth.2003.11.009","volume":"33","author":"O Gresch","year":"2004","unstructured":"Gresch, O. et al. New non-viral method for gene transfer into primary cells. Methods 33, 151\u2013163 (2004).","journal-title":"Methods"},{"key":"95848_CR21","doi-asserted-by":"publisher","first-page":"82","DOI":"10.1186\/1471-2121-11-82","volume":"11","author":"C Zhao","year":"2010","unstructured":"Zhao, C. et al. Spontaneously immortalised bovine mammary epithelial cells exhibits a distinct gene expression pattern from the breast cancer cells. BMC Cell. Biol. 11, 82 (2010).","journal-title":"BMC Cell. Biol."},{"key":"95848_CR22","first-page":"215","volume":"12","author":"R Harasawa","year":"1993","unstructured":"Harasawa, R., Hikiji, K., Tanabe, H., Takada, Y. & Mizusawa, H. Detection of adventitious Pestivirus in cell cultures by polymerase chain reaction using nested-pair primers. Tiss. Cult. Res. Commun. 12, 215\u2013220 (1993).","journal-title":"Tiss. Cult. Res. Commun."},{"key":"95848_CR23","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1007\/s10517-012-1648-1","volume":"156","author":"LV Uryvaev","year":"2012","unstructured":"Uryvaev, L. V. et al. Contamination of cell cultures with bovine viral diarrhea virus (BVDV). Bull. Exp. Biol. Med. 156, 77\u201381 (2012).","journal-title":"Bull. Exp. Biol. Med."},{"key":"95848_CR24","doi-asserted-by":"publisher","first-page":"e11165","DOI":"10.7717\/peerj.11165","volume":"9","author":"ZX Chin","year":"2021","unstructured":"Chin, Z. X. & Yeap, S. K. Ho WY Transfection types, methods and strategies: A technical review. PeerJ 9, e11165 (2021).","journal-title":"PeerJ"},{"key":"95848_CR25","doi-asserted-by":"publisher","first-page":"13","DOI":"10.2220\/biomedres.39.13","volume":"39","author":"N Toji","year":"2018","unstructured":"Toji, N. et al. A cell-based interferon-tau assay with an interferon-stimulated gene 15 promoter. Biomed. Res. 39, 13\u201320 (2018).","journal-title":"Biomed. Res."},{"key":"95848_CR26","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/j.vetmic.2007.11.004","volume":"129","author":"D Yamane","year":"2008","unstructured":"Yamane, D., Kato, K., Tohya, Y. & Akashi, H. The relationship between the viral RNA level and upregulation of innate immunity in spleen of cattle persistently infected with bovine viral diarrhea virus. Vet. Microbiol. 129, 69\u201379 (2008).","journal-title":"Vet. Microbiol."},{"key":"95848_CR27","doi-asserted-by":"publisher","first-page":"723","DOI":"10.1007\/s11259-009-9221-8","volume":"33","author":"C Liu","year":"2009","unstructured":"Liu, C., Chang, R., Yao, X., Qiao, W. T. & Geng, Y. Q. ISG15 expression in response to double-stranded RNA or LPS in cultured fetal bovine lung (FBL) cells. Vet. Res. Commun. 33, 723\u2013733 (2009).","journal-title":"Vet. Res. Commun."},{"key":"95848_CR28","doi-asserted-by":"publisher","first-page":"140","DOI":"10.1016\/j.jviromet.2006.08.005","volume":"138","author":"K Kameyama","year":"2006","unstructured":"Kameyama, K. et al. Development of an immunochromatographic test kit for rapid detection of bovine viral diarrhea virus antigen. J. Virol. Methods. 138, 140\u2013146 (2006).","journal-title":"J. Virol. Methods."},{"key":"95848_CR29","doi-asserted-by":"publisher","first-page":"480","DOI":"10.1007\/BF01863914","volume":"162","author":"G K\u00e4rber","year":"1931","unstructured":"K\u00e4rber, G. Beitrag zur kollektiven Behandlung pharmakologischer Reihenversuche Naunyn Schmiedebergs. Arch. Pharmacol. 162, 480\u2013483 (1931).","journal-title":"Arch. Pharmacol."},{"key":"95848_CR30","doi-asserted-by":"publisher","first-page":"452","DOI":"10.1038\/bmt.2012.244","volume":"48","author":"Y Kanda","year":"2013","unstructured":"Kanda, Y. Investigation of the freely available easy-to-use software \u2018EZR\u2019 for medical statistics. Bone Marrow Transplant. 48, 452\u2013458 (2013).","journal-title":"Bone Marrow Transplant."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-95848-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-95848-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-95848-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,3]],"date-time":"2022-12-03T12:16:22Z","timestamp":1670069782000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-021-95848-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,10]]},"references-count":30,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["95848"],"URL":"https:\/\/doi.org\/10.1038\/s41598-021-95848-0","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,8,10]]},"assertion":[{"value":"9 April 2021","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"30 July 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 August 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"16207"}}