{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T04:47:36Z","timestamp":1789620456224,"version":"build-2803163510"},"reference-count":47,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2022,1,7]],"date-time":"2022-01-07T00:00:00Z","timestamp":1641513600000},"content-version":"vor","delay-in-days":6,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Health Science Reports"],"published-print":{"date-parts":[[2022,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background and Aims<\/jats:title><jats:p>Autologous keratinocyte sheets constitute an important component of the burn wound treatment toolbox available to a surgeon and can be considered a life\u2010saving procedure for patients with severe burns over 50% of their total body surface area. Large\u2010scale keratinocyte sheet cultivation still fundamentally relies on the use of animal components such as inactivated murine 3T3 fibroblasts as feeders, animal\u2010derived enzymes such as trypsin, as well as media components such as fetal bovine serum (FBS). This study was therefore aimed to optimize autologous keratinocyte sheets by comparing various alternatives to critical components in their production.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Human skin samples were retrieved from remnant operative tissues. Cell isolation efficiency and viability were investigated by comparing the efficacy of porcine\u2010derived trypsin and animal\u2010free enzymes (Accutase and TrypLESelect). The subsequent expansion of the cells and the keratinocyte sheet formation was analyzed, comparing various cell culture substrates (inactivated murine 3T3 fibroblasts, inactivated human fibroblasts, Collagen I or plain tissue culture plastic), as well as media containing serum or chemically defined animal\u2010free media.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The cell isolation step showed clear cell yield advantages when using porcine\u2010derived trypsin, compared to animal\u2010free alternatives. The keratinocyte sheets produced using animal\u2010free serum were similar to those produced using 3T3 feeder layer and FBS\u2010containing medium, particularly in mechanical integrity as all grafts were liftable. In addition, sheets grown on collagen in an animal\u2010free medium showed indications of advantages in homogeneity, speed, reduced variability, and differentiation status compared to the other growth conditions investigated. Most importantly, the procedure was compatible with the up\u2010scaling requirements of major burn wound treatments.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This study demonstrated that animal\u2010free components could be used successfully to reduce the risk profile of large\u2010scale autologous keratinocyte sheet production, and thereby increase clinical accessibility.<\/jats:p><\/jats:sec>","DOI":"10.1002\/hsr2.449","type":"journal-article","created":{"date-parts":[[2022,1,7]],"date-time":"2022-01-07T09:24:22Z","timestamp":1641547462000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Optimizing large\u2010scale autologous human keratinocyte sheets for major burns\u2014Toward an animal\u2010free production and a more accessible clinical application"],"prefix":"10.1002","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2963-0420","authenticated-orcid":false,"given":"Laura","family":"Frese","sequence":"first","affiliation":[{"name":"Institute for Regenerative Medicine (IREM) University of Zurich  Zurich Switzerland"},{"name":"Center for Applied Biotechnology and Molecular Medicine (CABMM) University of Zurich  Zurich Switzerland"},{"name":"La Colline  Sion Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Salim Elias","family":"Darwiche","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology and Molecular Medicine (CABMM) University of Zurich  Zurich Switzerland"},{"name":"Musculoskeletal Research Unit (MSRU), Vetsuisse Faculty University of Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Myrna Elisabeth","family":"Gunning","sequence":"additional","affiliation":[{"name":"Department of Plastic and Reconstructive Surgery University Hospital Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simon Philipp","family":"Hoerstrup","sequence":"additional","affiliation":[{"name":"Institute for Regenerative Medicine (IREM) University of Zurich  Zurich Switzerland"},{"name":"Center for Applied Biotechnology and Molecular Medicine (CABMM) University of Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brigitte","family":"von Rechenberg","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology and Molecular Medicine (CABMM) University of Zurich  Zurich Switzerland"},{"name":"Musculoskeletal Research Unit (MSRU), Vetsuisse Faculty University of Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pietro","family":"Giovanoli","sequence":"additional","affiliation":[{"name":"Department of Plastic and Reconstructive Surgery University Hospital Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maurizio","family":"Calcagni","sequence":"additional","affiliation":[{"name":"Center for Applied Biotechnology and Molecular Medicine (CABMM) University of Zurich  Zurich Switzerland"},{"name":"Department of Plastic and Reconstructive Surgery University Hospital Zurich  Zurich Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,1,7]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM198408163110706"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0305-4179(96)00123-4"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1097\/BCR.0b013e3181e4ca29"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.burns.2006.11.002"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1186\/s41038-018-0117-0"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.metabol.2005.11.004"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.76.11.5665"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(75)80001-8"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0305-4179(99)00143-6"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.burns.2016.01.019"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/jbcr\/iry061"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.41.7.432"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1089\/152581601317210962"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.08-2115"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10561-011-9247-3"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02690-09"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.4061\/2011\/504723"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1186\/1475-925X-12-15"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10561-013-9368-y"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1186\/scrt8"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0806136105"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1067-1927.2004.12609.x"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-06934-3"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/exd.13328"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2009.241"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40472-018-0202-0"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-018-02986-5"},{"key":"e_1_2_10_29_1","unstructured":"PurtleDR CaffreyMB DoakJA EtchbergerKJ.Validated gamma irradiated trypsin powder"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0052494"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp060121o"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm1181"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biologicals.2011.08.003"},{"key":"e_1_2_10_34_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.11446"},{"key":"e_1_2_10_35_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0070263"},{"key":"e_1_2_10_36_1","doi-asserted-by":"publisher","DOI":"10.1155\/2016\/5246584"},{"key":"e_1_2_10_37_1","doi-asserted-by":"publisher","DOI":"10.1089\/ten.teb.2014.0547"},{"key":"e_1_2_10_38_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.040477"},{"key":"e_1_2_10_39_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00204-014-1250-6"},{"key":"e_1_2_10_40_1","doi-asserted-by":"publisher","DOI":"10.1634\/stemcells.2006-0444"},{"key":"e_1_2_10_41_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.TP.0000112381.80964.85"},{"key":"e_1_2_10_42_1","doi-asserted-by":"publisher","DOI":"10.1186\/s41038-017-0089-5"},{"key":"e_1_2_10_43_1","doi-asserted-by":"publisher","DOI":"10.5966\/sctm.2012-0072"},{"key":"e_1_2_10_44_1","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tec.2011.0255"},{"key":"e_1_2_10_45_1","doi-asserted-by":"publisher","DOI":"10.1002\/term.2118"},{"key":"e_1_2_10_46_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2020.00968"},{"key":"e_1_2_10_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiosc.2013.05.019"},{"key":"e_1_2_10_48_1","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tec.2014.0024"}],"container-title":["Health Science Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/hsr2.449","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/hsr2.449","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/hsr2.449","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T15:10:34Z","timestamp":1693235434000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/hsr2.449"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1]]},"references-count":47,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2022,1]]}},"alternative-id":["10.1002\/hsr2.449"],"URL":"https:\/\/doi.org\/10.1002\/hsr2.449","archive":["Portico"],"relation":{},"ISSN":["2398-8835","2398-8835"],"issn-type":[{"value":"2398-8835","type":"print"},{"value":"2398-8835","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1]]},"assertion":[{"value":"2021-02-11","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-10-25","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2022-01-07","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e449"}}