{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T09:01:40Z","timestamp":1777712500216,"version":"3.51.4"},"reference-count":47,"publisher":"SAGE Publications","issue":"4","license":[{"start":{"date-parts":[[2023,2,16]],"date-time":"2023-02-16T00:00:00Z","timestamp":1676505600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["The International Journal of Lower Extremity Wounds"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:p>\n                    <jats:bold>Background:<\/jats:bold>\n                    Off-the-shelf supply of viable engineered tissue is critical for effective and fast treatment of life-threatening injuries such as deep burns. An expanded keratinocyte sheet on the human amniotic membrane (KC sheet-HAM) is a beneficial tissue-engineering product for wound healing. To access an on-hand supply for the widespread application and overcome the time-consuming process, it is necessary to develop a cryopreservation protocol that guarantees the higher recovery of viable keratinocyte sheets after freeze-thawing. This research aimed to compare the recovery rate of KC sheet-HAM after cryopreservation by dimethyl-sulfoxide (DMSO) and glycerol.\n                    <jats:bold>Methods:<\/jats:bold>\n                    Amniotic membrane was decellularized with trypsin, and keratinocytes were cultured on it to form a multilayer, flexible, easy-to-handle KC sheet-HAM. The effects of 2 different cryoprotectants were investigated by histological analysis, live-dead staining, and proliferative capacity assessments before and after cryopreservation.\n                    <jats:bold>Results:<\/jats:bold>\n                    KCs well adhered and proliferated on the decellularized amniotic membrane and successfully represented 3 to 4 stratified layers of epithelialization after 2 to 3 weeks culture period; making it easy to cut, transfer, and cryopreserve. However, viability and proliferation assay indicated that both DMSO and glycerol cryosolutions have detrimental effects on KCs, and KCs-sheet HAM could not recover to the control level after 8 days of culture post-cryo. The KC sheet lost its stratified multilayer nature on AM, and sheet layers were reduced in both cryo-groups compared to the control.\n                    <jats:bold>Conclusion:<\/jats:bold>\n                    Expanding keratinocytes on the decellularized amniotic membrane as a multilayer sheet made a viable easy-to-handle sheet, nonetheless cryopreservation reduced viability and affected histological structure after thawing. Although some viable cells were detectable, our research highlighted the need for a better cryoprotectant protocol other than DMSO and glycerol, specific for the successful banking of viable tissue constructs.\n                  <\/jats:p>","DOI":"10.1177\/15347346231155751","type":"journal-article","created":{"date-parts":[[2023,2,16]],"date-time":"2023-02-16T04:08:31Z","timestamp":1676520511000},"page":"1119-1128","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":4,"title":["Comparing the Effects of Two Cryoprotectant Protocols, Dimethyl-Sulfoxide (DMSO) and Glycerol, on the Recovery Rate of Cultured Keratinocytes on Amniotic Membrane"],"prefix":"10.1177","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0642-1805","authenticated-orcid":false,"given":"Najmeh","family":"Mohammad-Pour","sequence":"first","affiliation":[{"name":"Department of Biology, Faculty of Science, Hakim Sabzevari University, Sabzevar, Iran"},{"name":"Culture and Research (ACECR)-Khorasan Razavi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1268-2224","authenticated-orcid":false,"given":"Vahid","family":"Moghimi","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science, Hakim Sabzevari University, Sabzevar, Iran"},{"name":"Culture and Research (ACECR)-Khorasan Razavi"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hamid Reza","family":"Bidkhori","sequence":"additional","affiliation":[{"name":"Culture and Research (ACECR)-Khorasan Razavi"},{"name":"Blood Borne Infections Research Center, Academic Center for Education, Culture and Research (ACECR)-Khorasan Razavi, Mashhad, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8372-1310","authenticated-orcid":false,"given":"Madjid","family":"Momeni-Moghaddam","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science, Hakim Sabzevari University, Sabzevar, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hojjat","family":"Naderi-Meshkin","sequence":"additional","affiliation":[{"name":"Culture and Research (ACECR)-Khorasan Razavi"},{"name":"Wellcome-Wolfson Institute for Experimental Medicine, Belfast, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2023,2,16]]},"reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(75)90183-X"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(81)90006-4"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2013.02.001"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1524-4725.2012.02561.x"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1742-481X.2012.01083.x"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1001\/archderm.1992.01680150063007"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1590\/S0102-86502013001200004"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.reth.2017.03.002"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1001\/archderm.1993.01680280067013"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1524-4725.1999.99022.x"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0305-4179(01)00008-0"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1089\/biores.2016.0036"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1089\/ten.tec.2014.0600"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1002\/term.1690"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.burns.2006.11.002"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1177\/1534734619859214"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jcyt.2016.10.008"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10616-015-9940-3"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a015156"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/bmb\/ldv053"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-012-1424-6"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0161-6420(02)01105-3"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.survophthal.2003.08.003"},{"issue":"1","key":"e_1_3_2_25_2","first-page":"18","article-title":"Reconstruction of damaged corneas by transplantation of autologous limbal epithelial cells","volume":"2","author":"Tsai RJF","year":"2001","unstructured":"Tsai RJF, Li L, Chen J, Ladas JG, Stark W. 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