{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T11:23:15Z","timestamp":1772104995084,"version":"3.50.1"},"reference-count":38,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T00:00:00Z","timestamp":1771891200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Purpose: To establish and comprehensively characterize immortalized limbal epithelial cell lines derived from patients with PAX6 haploinsufficiency-associated congenital aniridia, as well as from a healthy donor. These well-defined cell models provide a reliable and reproducible platform for long-term experimental studies, facilitating mechanistic investigations and the development and evaluation of novel therapeutic approaches. Methods: Primary limbal epithelial cells were isolated from biopsies of two patients with distinct PAX6 variants and from a healthy donor. Immortalization was performed by InSCREENex GmbH. The resulting cell lines were characterized using microscopy, BrdU assay, qPCR, flow cytometry, immunocytochemistry, and mRNA sequencing. Results: Immortalized aniridia and control cell lines displayed typical polygonal epithelial morphology and comparable proliferation rates. Total PAX6 mRNA and protein levels were similar among groups; however, nuclear PAX6 immunosignals were significantly reduced in aniridia-derived lines. Expression of ABCG2, TP63, FOSL2, ALDH1A1, and FABP5 showed no significant differences, except for reduced \u0394Np63\u03b1 protein levels in one aniridia line. mRNA sequencing detected more than 14,000 transcripts, including subsets uniquely expressed in control and aniridia-derived lines. Conclusions: Immortalized aniridia limbal epithelial cell lines preserve key epithelial characteristics and overall transcriptomic similarity to controls while exhibiting disease-relevant molecular alterations. These cell lines represent models of PAX6-associated ocular surface disease.<\/jats:p>","DOI":"10.3390\/cells15050394","type":"journal-article","created":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T15:05:27Z","timestamp":1771945527000},"page":"394","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Patient-Derived Immortalized Limbal Epithelial Cells as In Vitro Models of Congenital Aniridia"],"prefix":"10.3390","volume":"15","author":[{"given":"Tanja","family":"Stachon","sequence":"first","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Experimental Ophthalmology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shweta","family":"Suiwal","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Experimental Ophthalmology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Virendra","family":"Kumar","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tobias","family":"May","sequence":"additional","affiliation":[{"name":"InSCREENex GmbH, 38124 Braunschweig, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frank","family":"Schmitz","sequence":"additional","affiliation":[{"name":"Institute of Anatomy and Cell Biology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maryam","family":"Amini","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8526-4712","authenticated-orcid":false,"given":"Fabian N.","family":"Fries","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Ophthalmology, Saarland University Medical Center, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9701-8204","authenticated-orcid":false,"given":"Berthold","family":"Seitz","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Saarland University Medical Center, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5235-5039","authenticated-orcid":false,"given":"Shao-Lun","family":"Hsu","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Experimental Ophthalmology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-2251-6242","authenticated-orcid":false,"given":"Shuailin","family":"Li","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Experimental Ophthalmology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shanhe","family":"Liu","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Experimental Ophthalmology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Swarnali","family":"Kundu","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole","family":"Ludwig","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Center for Human and Molecular Biology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N\u00f3ra","family":"Szentm\u00e1ry","sequence":"additional","affiliation":[{"name":"Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Aniridia Research, Saarland University, 66421 Saarbr\u00fccken, Germany"},{"name":"Department of Experimental Ophthalmology, Saarland University, 66421 Saarbr\u00fccken, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.jtos.2021.09.001","article-title":"Pathophysiology of Aniridia-Associated Keratopathy: Developmental Aspects and Unanswered Questions","volume":"22","author":"Latta","year":"2021","journal-title":"Ocul. Surf."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.survophthal.2017.09.004","article-title":"The Genetics of Congenital Aniridia\u2014A Guide for the Ophthalmologist","volume":"63","author":"Landsend","year":"2018","journal-title":"Surv. 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