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Long-Term Cultures of Human Cornea Limbal Explants Form 3D Structures Ex Vivo – Implications for Tissue Engineering and Clinical Applications

Ljubimov, Alexander V., Szabó, Dóra Júlia, Noer, Agate, Nagymihály, Richárd, Josifovska, Natasha, Andjelic, Sofija, Veréb, Zoltán, Facskó, Andrea, Moe, Morten C. and Petrovski, Goran (2015) Long-Term Cultures of Human Cornea Limbal Explants Form 3D Structures Ex Vivo – Implications for Tissue Engineering and Clinical Applications. PLOS ONE, 10 (11). e0143053. [Journal article]

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URL: http://dx.doi.org/10.1371/journal.pone.0143053

DOI: 10.1371/journal.pone.0143053

Abstract

Long-term cultures of cornea limbal epithelial stem cells (LESCs) were developed and characterized for future tissue engineering and clinical applications. The limbal tissue explants were cultivated and expanded for more than 3 months in medium containing serum as the only growth supplement and without use of scaffolds. Viable 3D cell outgrowth from the explants was observed within 4 weeks of cultivation. The outgrowing cells were examined by immunofluorescent staining for putative markers of stemness (ABCG2, CK15, CK19 and Vimentin), proliferation (p63α, Ki-67), limbal basal epithelial cells (CK8/18) and differentiated cornea epithelial cells (CK3 and CK12). Morphological and immunostaining analyses revealed that long-term culturing can form stratified 3D tissue layers with a clear extracellular matrix deposition and organization (collagen I, IV and V). The LESCs showed robust expression of p63α, ABCG2, and their surface marker fingerprint (CD117/c-kit, CXCR4, CD146/MCAM, CD166/ALCAM) changed over time compared to short-term LESC cultures. Overall, we provide a model for generating stem cell-rich, long-standing 3D cultures from LESCs which can be used for further research purposes and clinical transplantation

Item Type:Journal article
Keywords:cornea, limbal epithelial stem cells, long-term cultures, 3D tissue explants
Faculties and Schools:Faculty of Life and Health Sciences > School of Biomedical Sciences
Faculty of Life and Health Sciences
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Optometry and Vision Science
ID Code:38148
Deposited By: Dr Natasha Josifovska
Deposited On:13 Jun 2017 09:26
Last Modified:13 Jun 2017 09:26

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