{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,19]],"date-time":"2025-09-19T09:37:49Z","timestamp":1758274669142,"version":"3.41.2"},"reference-count":27,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2022,1,27]],"date-time":"2022-01-27T00:00:00Z","timestamp":1643241600000},"content-version":"vor","delay-in-days":26,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2022,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Efficient gene transfer into cultured fibroblasts and keratinocytes during retroviral transduction is a critical step toward the treatment of genodermatoses such as epidermolysis bullosa. However, achieving high transduction rates is still a difficult task, particularly for the insertion of large coding sequences for which high viral titers cannot always be obtained. Multiple polycationic molecules, such as polybrene, which has been used in several clinical trials, have the ability to boost ex vivo retroviral gene transfer. However, the use of polybrene has been associated with a reduction of the proliferation and growth potential of human keratinocytes in culture. We developed a method for the efficient retroviral transduction of primary fibroblasts and keratinocytes using EF\u2010c, a polycationic nanofibril\u2010forming peptide. In comparison with polybrene, we found that the retroviral transduction efficiency with EF\u2010c was increased 2.5\u2010 to 3.2\u2010fold for fibroblasts, but not for keratinocytes. Moreover, the use of EF\u2010c did not affect fibroblast proliferation and keratinocyte stem cell content, whereas polybrene induced a decrease in both. This method could have a positive impact on the development of ex vivo gene correction of genodermatoses, allowing for more efficient gene transfer into primary skin cells with little to no effect on proliferation and stem cell content. \u00a9 2022 Wiley Periodicals LLC.<\/jats:p><jats:p><jats:bold>Basic Protocol<\/jats:bold>: Fibroblast and keratinocyte transduction<\/jats:p><jats:p><jats:bold>Support Protocol<\/jats:bold>: Assessment of transduction efficiency through flow cytometry analysis<\/jats:p>","DOI":"10.1002\/cpz1.353","type":"journal-article","created":{"date-parts":[[2022,1,27]],"date-time":"2022-01-27T20:07:00Z","timestamp":1643314020000},"source":"Crossref","is-referenced-by-count":2,"title":["Efficient Gamma\u2010Retroviral Transduction of Primary Human Skin Cells Using the EF\u2010c Peptide as a Transduction Enhancer"],"prefix":"10.1002","volume":"2","author":[{"given":"Martin A.","family":"Barbier","sequence":"first","affiliation":[{"name":"Centre de recherche en organog\u00e9n\u00e8se exp\u00e9rimentale de l'Universit\u00e9 Laval\/LOEX et D\u00e9partement de chirurgie, Facult\u00e9 de m\u00e9decine Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"},{"name":"Centre de recherche du CHU de Qu\u00e9bec\u2010Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela Dakiw","family":"Piaceski","sequence":"additional","affiliation":[{"name":"Centre de recherche en organog\u00e9n\u00e8se exp\u00e9rimentale de l'Universit\u00e9 Laval\/LOEX et D\u00e9partement de chirurgie, Facult\u00e9 de m\u00e9decine Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"},{"name":"Centre de recherche du CHU de Qu\u00e9bec\u2010Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Danielle","family":"Larouche","sequence":"additional","affiliation":[{"name":"Centre de recherche en organog\u00e9n\u00e8se exp\u00e9rimentale de l'Universit\u00e9 Laval\/LOEX et D\u00e9partement de chirurgie, Facult\u00e9 de m\u00e9decine Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"},{"name":"Centre de recherche du CHU de Qu\u00e9bec\u2010Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah H.","family":"Villeneuve","sequence":"additional","affiliation":[{"name":"Centre de recherche en organog\u00e9n\u00e8se exp\u00e9rimentale de l'Universit\u00e9 Laval\/LOEX et D\u00e9partement de chirurgie, Facult\u00e9 de m\u00e9decine Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"},{"name":"Centre de recherche du CHU de Qu\u00e9bec\u2010Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Karim","family":"Ghani","sequence":"additional","affiliation":[{"name":"Centre de recherche du CHU de Qu\u00e9bec\u2010Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"},{"name":"Centre de recherche sur le cancer de l'Universit\u00e9 Laval, D\u00e9partement de biologie mol\u00e9culaire, de biochimie m\u00e9dicale et de pathologie, Facult\u00e9 de m\u00e9decine Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elena","family":"Pope","sequence":"additional","affiliation":[{"name":"Hospital for Sick Children University of Toronto  Toronto Ontario Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manuel","family":"Caruso","sequence":"additional","affiliation":[{"name":"Centre de recherche du CHU de Qu\u00e9bec\u2010Universit\u00e9 Laval  Qu\u00e9bec City Quebec Canada"},{"name":"Centre de recherche sur le cancer de l'Universit\u00e9 Laval, D\u00e9partement de biologie mol\u00e9culaire, de biochimie m\u00e9dicale et de pathologie, Facult\u00e9 de m\u00e9decine Universit\u00e9 Laval  Qu\u00e9bec City Quebec 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