{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T06:10:33Z","timestamp":1785391833738,"version":"3.55.0"},"reference-count":35,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2023,5,10]],"date-time":"2023-05-10T00:00:00Z","timestamp":1683676800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>In this study we describe peripheral corneal endothelial cell migration<jats:italic>in vitro<\/jats:italic>in the absence and presence of a ROCK-inhibitor. For this study, 21 corneal endothelial graft rims, with attached trabecular meshwork (TM), were prepared from Descemet membrane-endothelial cell sheets by 6.5 mm trepanation. For the initial proof-of-concept, 7 outer graft rims were cultured in a thermo-reversible hydrogel matrix for up to 47 days. To assess the effect of a ROCK-inhibitor, 14 paired outer rims were cultured either with or without ROCK-inhibitor for up to 46 days. At the end of culture, tissue was retrieved from the hydrogel matrix and examined for cell viability and expression of different endothelial cell markers (ZO-1, Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>-ATPase, NCAM, glypican, and vimentin). All cultured rims remained viable and displayed either single regions (n = 5\/21) or collective areas (n = 16\/21) of cell migration, regardless of the presence or absence of ROCK-inhibition. Migration started after 4\u00b12 days and continued for at least 29 days. The presence of ROCK-inhibitor seemed to contribute to a more regular cell morphology of migrating cells. In addition, 7 outer rims demonstrated a phenotypically distinct late-onset but fast-growing cell population emerging from the area close to the limbus. These cells emerged after 3 weeks of culture and appeared less differentiated compared to other areas of migration. Immunostaining showed that migrated cells maintained the expression patterns of endothelial cell markers. In conclusion, we observed 2 morphologically distinct migrating cell populations with the first type being triggered by a broken physical barrier, which disrupted contact inhibition and the second, late-onset type showing a higher proliferative capacity though appearing less differentiated. This cell subpopulation appeared to be mediated by stimuli other than loss of contact inhibition and ROCK-inhibitor presence. Further exploration of the differences between these cell types may assist in optimizing regenerative treatment options for endothelial diseases.<\/jats:p>","DOI":"10.1371\/journal.pone.0285609","type":"journal-article","created":{"date-parts":[[2023,5,10]],"date-time":"2023-05-10T18:33:04Z","timestamp":1683743584000},"page":"e0285609","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":14,"title":["Early and late-onset cell migration from peripheral corneal endothelium"],"prefix":"10.1371","volume":"18","author":[{"given":"Alina","family":"Miron","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sorcha","family":"N\u00ed Dhubhghaill","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3044-5057","authenticated-orcid":true,"given":"Viridiana","family":"Kocaba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martine J.","family":"Jager","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5067-3834","authenticated-orcid":true,"given":"Gerrit R. J.","family":"Melles","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5703-5971","authenticated-orcid":true,"given":"Silke","family":"Oellerich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,5,10]]},"reference":[{"key":"pone.0285609.ref001","doi-asserted-by":"crossref","first-page":"107794","DOI":"10.1016\/j.exer.2019.107794","article-title":"Experimental models of corneal endothelial cell therapy and translational challenges to clinical practice","volume":"188","author":"K Rolev","year":"2019","journal-title":"Exp Eye Res"},{"key":"pone.0285609.ref002","doi-asserted-by":"crossref","first-page":"108462","DOI":"10.1016\/j.exer.2021.108462","article-title":"Focus on cell therapy to treat corneal endothelial diseases","volume":"204","author":"PA Faye","year":"2021","journal-title":"Exp Eye Res"},{"key":"pone.0285609.ref003","doi-asserted-by":"crossref","first-page":"100987","DOI":"10.1016\/j.preteyeres.2021.100987","article-title":"Approaches for corneal endothelium regenerative medicine","volume":"87","author":"P Catal\u00e0","year":"2022","journal-title":"Prog Retin Eye Res"},{"key":"pone.0285609.ref004","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.survophthal.2017.07.004","article-title":"A review of the evidence for in vivo corneal endothelial regeneration","volume":"63","author":"B Van den Bogerd","year":"2018","journal-title":"Surv Ophthalmol"},{"key":"pone.0285609.ref005","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.trsl.2022.05.003","article-title":"Corneal endothelial wound healing: understanding the regenerative capacity of the innermost layer of the cornea","volume":"248","author":"H Vercammen","year":"2022","journal-title":"Transl Res"},{"key":"pone.0285609.ref006","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.jcjo.2014.10.014","article-title":"Descemetorhexis for Fuchs\u2019 dystrophy","volume":"50","author":"G Moloney","year":"2015","journal-title":"Can J Ophthalmol"},{"key":"pone.0285609.ref007","doi-asserted-by":"crossref","first-page":"1267","DOI":"10.1097\/ICO.0000000000000915","article-title":"Treatment of Fuchs endothelial dystrophy by Descemet stripping without endothelial keratoplasty","volume":"35","author":"DS Borkar","year":"2016","journal-title":"Cornea"},{"key":"pone.0285609.ref008","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1097\/ICO.0000000000001202","article-title":"Descemetorhexis without graft placement for the treatment of Fuchs endothelial dystrophy: preliminary results and review of the literature","volume":"36","author":"A Iovieno","year":"2017","journal-title":"Cornea"},{"key":"pone.0285609.ref009","doi-asserted-by":"crossref","first-page":"1225","DOI":"10.1097\/ICO.0000000000002642","article-title":"A close look at the clinical efficacy of Rho-associated protein kinase inhibitor eye drops for Fuchs endothelial corneal dystrophy","volume":"40","author":"S Kinoshita","year":"2021","journal-title":"Cornea"},{"key":"pone.0285609.ref010","doi-asserted-by":"crossref","first-page":"e0138076","DOI":"10.1371\/journal.pone.0138076","article-title":"Recovery of corneal endothelial cells from periphery after injury","volume":"10","author":"SO Choi","year":"2015","journal-title":"PLoS One."},{"key":"pone.0285609.ref011","doi-asserted-by":"crossref","first-page":"2523","DOI":"10.1002\/stem.1212","article-title":"Revisited microanatomy of the corneal endothelial periphery: new evidence for continuous centripetal migration of endothelial cells in humans","volume":"30","author":"Z He","year":"2012","journal-title":"Stem Cells"},{"key":"pone.0285609.ref012","doi-asserted-by":"crossref","first-page":"2190","DOI":"10.1089\/scd.2013.0387","article-title":"Identification and potential application of human corneal endothelial progenitor cells","volume":"23","author":"S Hara","year":"2014","journal-title":"Stem Cells Dev"},{"key":"pone.0285609.ref013","doi-asserted-by":"crossref","first-page":"1244","DOI":"10.3390\/cells8101244","article-title":"Characterization of human transition zone reveals a putative progenitor-enriched niche of corneal endothelium","volume":"8","author":"GHF Yam","year":"2019","journal-title":"Cells"},{"key":"pone.0285609.ref014","first-page":"412743","article-title":"Progenitors for the corneal endothelium and trabecular meshwork: a potential source for personalized stem cell therapy in corneal endothelial diseases and glaucoma.","author":"WY Yu","year":"2011","journal-title":"J Biomed Biotechnol"},{"key":"pone.0285609.ref015","doi-asserted-by":"crossref","first-page":"642","DOI":"10.1097\/ICO.0000000000001209","article-title":"Descemetorhexis without grafting for Fuchs endothelial dystrophy-supplementation with topical Ripasudil","volume":"36","author":"G Moloney","year":"2017","journal-title":"Cornea"},{"key":"pone.0285609.ref016","doi-asserted-by":"crossref","first-page":"1479","DOI":"10.1097\/ICO.0000000000001742","article-title":"Descemetorhexis without endothelial keratoplasty versus DMEK for treatment of Fuchs endothelial corneal dystrophy","volume":"37","author":"MJ Huang","year":"2018","journal-title":"Cornea"},{"key":"pone.0285609.ref017","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1097\/ICO.0000000000001883","article-title":"Use of topical Rho kinase inhibitors in the treatment of Fuchs dystrophy after Descemet stripping only","volume":"38","author":"MS Macsai","year":"2019","journal-title":"Cornea"},{"key":"pone.0285609.ref018","doi-asserted-by":"crossref","first-page":"1048","DOI":"10.1097\/ICO.0000000000002270","article-title":"5-Year outcomes of Descemet stripping only in Fuchs dystrophy","volume":"39","author":"JA Artieda","year":"2020","journal-title":"Cornea"},{"key":"pone.0285609.ref019","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1097\/ICO.0000000000002691","article-title":"Pilot Study of corneal clearance with the use of a Rho-kinase inhibitor after descemetorhexis without endothelial keratoplasty for Fuchs endothelial corneal dystrophy","volume":"40","author":"E Davies","year":"2021","journal-title":"Cornea"},{"key":"pone.0285609.ref020","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1097\/ICU.0000000000000579","article-title":"Descemet\u2019s stripping without endothelial keratoplasty","volume":"30","author":"D Garcerant","year":"2019","journal-title":"Curr Opin Ophthalmol"},{"key":"pone.0285609.ref021","doi-asserted-by":"crossref","first-page":"1578","DOI":"10.1016\/j.jcrs.2008.05.036","article-title":"Donor tissue preparation for Descemet membrane endothelial keratoplasty","volume":"34","author":"JT Lie","year":"2008","journal-title":"J Cataract Refract Surg"},{"key":"pone.0285609.ref022","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1111\/j.1755-3768.2012.02462.x","article-title":"Standardized \u2018no-touch\u2019 donor tissue preparation for DALK and DMEK: harvesting undamaged anterior and posterior transplants from the same donor cornea.","volume":"91","author":"EA Groeneveld-van Beek","year":"2013","journal-title":"Acta Ophthalmol"},{"key":"pone.0285609.ref023","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1080\/02713683.2020.1798468","article-title":"Improving endothelial explant tissue culture by novel thermoresponsive cell culture system","volume":"46","author":"A Miron","year":"2021","journal-title":"Curr Eye Res"},{"key":"pone.0285609.ref024","doi-asserted-by":"crossref","first-page":"e0225462","DOI":"10.1371\/journal.pone.0225462","article-title":"In vitro endothelial cell migration from limbal edge-modified Quarter-DMEK grafts","volume":"14","author":"A Miron","year":"2019","journal-title":"PLoS One"},{"key":"pone.0285609.ref025","doi-asserted-by":"crossref","first-page":"e0246516","DOI":"10.1371\/journal.pone.0246516","article-title":"Preclinical testing of small diameter Descemet membrane endothelial keratoplasty grafts to increase tissue availability","volume":"16","author":"SN Dhubhghaill","year":"2021","journal-title":"PLoS One"},{"key":"pone.0285609.ref026","doi-asserted-by":"crossref","first-page":"8097","DOI":"10.1038\/s41598-017-08232-2","article-title":"Direct reprogramming of fibroblasts into skeletal muscle progenitor cells by transcription factors enriched in undifferentiated subpopulation of satellite cells","volume":"7","author":"N Ito","year":"2017","journal-title":"Sci Rep"},{"key":"pone.0285609.ref027","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1186\/s13287-020-02046-2","article-title":"Regenerative capacity of the corneal transition zone for endothelial cell therapy","volume":"11","author":"NM Ming","year":"2020","journal-title":"Stem Cell Res Ther"},{"key":"pone.0285609.ref028","doi-asserted-by":"crossref","first-page":"871","DOI":"10.2217\/rme-2020-0202","article-title":"Challenges in corneal endothelial cell culture","volume":"16","author":"R Wongvisavavit","year":"2021","journal-title":"Regen Med"},{"key":"pone.0285609.ref029","doi-asserted-by":"crossref","first-page":"1626","DOI":"10.1167\/iovs.04-1263","article-title":"Human corneal endothelial cell precursors isolated by sphere-forming assay","volume":"46","author":"S Yokoo","year":"2005","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0285609.ref030","doi-asserted-by":"crossref","first-page":"3645","DOI":"10.1167\/iovs.05-0630","article-title":"Comparison of rabbit corneal endothelial cell precursors in the central and peripheral cornea","volume":"46","author":"T Mimura","year":"2005","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0285609.ref031","doi-asserted-by":"crossref","first-page":"3128","DOI":"10.1167\/iovs.05-0251","article-title":"Sphere therapy for corneal endothelium deficiency in a rabbit model","volume":"46","author":"T Mimura","year":"2005","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0285609.ref032","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1111\/ceo.13756","article-title":"Successful culture of human transition zone cells","volume":"48","author":"J Zhang","year":"2020","journal-title":"Clin Exp Ophthalmol"},{"key":"pone.0285609.ref033","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s12929-018-0492-7","article-title":"Advances in culture, expansion and mechanistic studies of corneal endothelial cells: a systematic review","volume":"26","author":"S Chen","year":"2019","journal-title":"J Biomed Sci"},{"key":"pone.0285609.ref034","doi-asserted-by":"crossref","first-page":"e62095","DOI":"10.1371\/journal.pone.0062095","article-title":"ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells","volume":"8","author":"A Pipparelli","year":"2013","journal-title":"PLoS One"},{"key":"pone.0285609.ref035","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.3892\/ijmm.2017.3103","article-title":"The ROCK inhibitor, thiazovivin, inhibits human corneal endothelial\u2011to\u2011mesenchymal transition\/epithelial\u2011to\u2011mesenchymal transition and increases ionic transporter expression","volume":"40","author":"Q Wu","year":"2017","journal-title":"Int J Mol Med"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0285609","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,12,12]],"date-time":"2023-12-12T15:21:01Z","timestamp":1702394461000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0285609"}},"subtitle":[],"editor":[{"given":"Yu-Chi","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2023,5,10]]},"references-count":35,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,5,10]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0285609","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,5,10]]}}}