{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T11:20:09Z","timestamp":1761218409718,"version":"build-2065373602"},"reference-count":16,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,8,16]],"date-time":"2021-08-16T00:00:00Z","timestamp":1629072000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e a Tecnologia","doi-asserted-by":"publisher","award":["SFRH\/BD\/137067\/2018","UIDB\/00276\/2020"],"award-info":[{"award-number":["SFRH\/BD\/137067\/2018","UIDB\/00276\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Commercial hare and rabbit immortalized cell lines are extremely limited regarding the many species within the lagomorpha order. To overcome this limitation, researchers and technicians must establish primary cell cultures derived from biopsies or embryos. Among all cell types, fibroblasts are plastic and resilient cells, highly convenient for clinical and fundamental research but also for diagnosis, particularly for viral isolation. Here, we describe a fast and cheap method to produce primary fibroblast cell cultures from leporid species, using dispase II, a protease that allows dermal\u2013epidermal separation, followed by a simple enzymatic digestion with trypsin. This method allows for the establishment of an in vitro cell culture system with an excellent viability yield and purity level higher than 85% and enables the maintenance and even immortalization of leporid fibroblastic cells derived from tissues already differentiated.<\/jats:p>","DOI":"10.3390\/cells10082100","type":"journal-article","created":{"date-parts":[[2021,8,16]],"date-time":"2021-08-16T21:27:45Z","timestamp":1629149265000},"page":"2100","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Simple Method for Establishing Primary Leporidae Skin Fibroblast Cultures"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0696-7322","authenticated-orcid":false,"given":"F\u00e1bio A.","family":"Abade dos Santos","sequence":"first","affiliation":[{"name":"Centro de Investiga\u00e7\u00e3o Interdisciplinar em Sanidade Animal (CIISA), Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa, Avenida da Universidade T\u00e9cnica, 1300-477 Lisboa, Portugal"},{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"},{"name":"Instituto Universitario de Biotecnolog\u00eda de Asturias (IUBA), Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular, Universidad de Oviedo, 33003 Oviedo, Spain"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6648-9254","authenticated-orcid":false,"given":"C. L.","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"given":"Isabel","family":"Almeida","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"given":"Teresa","family":"Fagulha","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"given":"Fernanda","family":"Rammos","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6246-1983","authenticated-orcid":false,"given":"S\u00edlvia C.","family":"Barros","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"given":"Margarida","family":"Henriques","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8527-3731","authenticated-orcid":false,"given":"Tiago","family":"Lu\u00eds","sequence":"additional","affiliation":[{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1488-9659","authenticated-orcid":false,"given":"Margarida D.","family":"Duarte","sequence":"additional","affiliation":[{"name":"Centro de Investiga\u00e7\u00e3o Interdisciplinar em Sanidade Animal (CIISA), Faculdade de Medicina Veterin\u00e1ria, Universidade de Lisboa, Avenida da Universidade T\u00e9cnica, 1300-477 Lisboa, Portugal"},{"name":"Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV, I.P.), Av. da Rep\u00fablica, Quinta do Marqu\u00eas, 2780-157 Oeiras, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13578-018-0264-9","article-title":"A prospect of cell immortalization combined with matrix microenvironmental optimization strategy for tissue engineering and regeneration","volume":"9","author":"Wang","year":"2019","journal-title":"Cell Biosci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-019-39770-6","article-title":"Fibroblast origin shapes tissue homeostasis, epidermal differentiation, and drug uptake","volume":"9","author":"Hausmann","year":"2019","journal-title":"Sci. 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