{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:49:40Z","timestamp":1792788580693,"version":"4.5.0"},"reference-count":33,"publisher":"The Company of Biologists","issue":"17","license":[{"start":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T00:00:00Z","timestamp":1694476800000},"content-version":"vor","delay-in-days":11,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000072","name":"National Institute of Dental and Craniofacial Research","doi-asserted-by":"crossref","award":["R01DE022087"],"award-info":[{"award-number":["R01DE022087"]}],"id":[{"id":"10.13039\/100000072","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Ohio State University","award":["P30CA016058"],"award-info":[{"award-number":["P30CA016058"]}]},{"name":"National Cancer Institute"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,9,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Conditional reprogramming is a cell culture technique that effectively immortalizes epithelial cells with normal genotypes by renewing epidermal stem cells. Y-27632, a compound that promotes conditional reprogramming through an unknown mechanism, was developed to inhibit the two Rho-associated kinase (ROCK) isoforms. We used human foreskin keratinocytes (HFKs) to study the role of Y-27632 in conditional reprogramming and learn how ROCKs control epidermal stem cell renewal. In conditional reprogramming, Y-27632 increased HFK adherence to culture dishes, progression through S, G2 and M phases of the cell cycle, and epidermal stem cell marker levels. Although this correlated with ROCK inhibition by Y-27632, we generated CRISPR\u2013Cas9-mediated HFK ROCK knockouts to test the direct role of ROCK inhibition. Knockout of single ROCK isoforms was insufficient to disrupt ROCK activity or promote HFK propagation without Y-27632. Although ROCK activity was reduced, HFKs with double knockout of ROCK1 and ROCK2 still required Y-27632 to propagate. Y-27632 was the most effective among the ROCK inhibitors we tested at promoting HFK proliferation and epidermal stem cell marker expression. Thus, the ability of Y-27632 to promote an epidermal stem cell state in conditional reprogramming not only depends upon ROCK inhibition but also acts via as-yet-unidentified mechanisms. Epidermal stem cell renewal might in part be regulated by ROCKs, but also involves additional pathways.<\/jats:p>","DOI":"10.1242\/jcs.260990","type":"journal-article","created":{"date-parts":[[2023,8,10]],"date-time":"2023-08-10T08:03:43Z","timestamp":1691654623000},"source":"Crossref","is-referenced-by-count":16,"title":["Y-27632 acts beyond ROCK inhibition to maintain epidermal stem-like cells in culture"],"prefix":"10.1242","volume":"136","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2695-8148","authenticated-orcid":false,"given":"Travis A.","family":"Witkowski","sequence":"first","affiliation":[{"name":"The Ohio State University Wexner Medical Center 1 Department of Otolaryngology \u2013 Head and Neck Surgery , , Columbus, OH 43210 , USA"},{"name":"The James Cancer Hospital and Solove Research Institute, The Ohio State University 2 , Columbus, OH 43210 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Li","sequence":"additional","affiliation":[{"name":"The Ohio State University Wexner Medical Center 1 Department of Otolaryngology \u2013 Head and Neck Surgery , , Columbus, OH 43210 , USA"},{"name":"The James Cancer Hospital and Solove Research Institute, The Ohio State University 2 , Columbus, OH 43210 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason G.","family":"Andersen","sequence":"additional","affiliation":[{"name":"The Ohio State University Wexner Medical Center 1 Department of Otolaryngology \u2013 Head and Neck Surgery , , Columbus, OH 43210 , USA"},{"name":"The James Cancer Hospital and Solove Research Institute, The Ohio State University 2 , Columbus, OH 43210 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bhavna","family":"Kumar","sequence":"additional","affiliation":[{"name":"The Ohio State University Wexner Medical Center 1 Department of Otolaryngology \u2013 Head and Neck Surgery , , Columbus, OH 43210 , USA"},{"name":"The James Cancer Hospital and Solove Research Institute, The Ohio State University 2 , Columbus, OH 43210 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1014-7853","authenticated-orcid":false,"given":"Edmund A.","family":"Mroz","sequence":"additional","affiliation":[{"name":"The Ohio State University Wexner Medical Center 1 Department of Otolaryngology \u2013 Head and Neck Surgery , , Columbus, OH 43210 , USA"},{"name":"The James Cancer Hospital and Solove Research Institute, The Ohio State University 2 , Columbus, OH 43210 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5312-0303","authenticated-orcid":false,"given":"James W.","family":"Rocco","sequence":"additional","affiliation":[{"name":"The Ohio State University Wexner Medical Center 1 Department of Otolaryngology \u2013 Head and Neck Surgery , , Columbus, OH 43210 , USA"},{"name":"The James Cancer Hospital and Solove Research Institute, The Ohio State University 2 , Columbus, OH 43210 , USA"},{"name":"The Ohio State University Comprehensive Cancer Center \u2013 James, The Ohio State University 3 , Columbus, OH 43210 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2023,9,12]]},"reference":[{"key":"2024061307500849300_JCS260990C1","doi-asserted-by":"publisher","first-page":"371","DOI":"10.1038\/nprot.2009.241","article-title":"Isolation and cultivation of human keratinocytes from skin or plucked hair for the generation of induced pluripotent stem cells","volume":"5","author":"Aasen","year":"2010","journal-title":"Nat. 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