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

Alexander V. Ljubimov, Dóra Júlia Szabó, Agate Noer, Richárd Nagymihály, Natasha Josifovska, Sofija Andjelic, Zoltán Veréb, Andrea Facskó, Morten C. Moe, Goran Petrovski

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    17 Citations (Scopus)

    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
    Original languageEnglish
    Pages (from-to)e0143053
    JournalPLoS ONE
    Volume10
    Issue number11
    DOIs
    Publication statusPublished - 18 Nov 2015

    Keywords

    • cornea
    • limbal epithelial stem cells
    • long-term cultures
    • 3D tissue explants

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  • Cite this

    Ljubimov, A. V., Szabó, D. J., Noer, A., Nagymihály, R., Josifovska, N., Andjelic, S., Veréb, Z., Facskó, A., Moe, M. C., & Petrovski, G. (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. https://doi.org/10.1371/journal.pone.0143053