Purchase this article with an account.
Goran Petrovski, Dora Julia Szabo, Richard Nagymihaly, Natasha Josifovska, Sofija Andjelic, Zoltan Janos Vereb, Andrea Facsko, Morten Carstens Moe, Agate Noer; Long-standing cultures from human cornea limbal and stromal grafts form 3D structures ex vivo - potential implications for future transplantation. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3452.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Long-standing cultivation and characterizion of human cornea limbal epithelial- and stromal stem cells (LESCs and CSSCs), respectively, into 3D tissue grafts was developed for future tissue engineering and clinical applications.
The limbal and stromal tissue explants were obtained from cadavers according to the guidelines of the Helsinki Declaration, and cultivated and expanded for more than 3 months in medium containing serum as the only growth supplement. Viable 3D cell outgrowth from the explants was observed within 4 weeks of cultivation. The outgrowing cells were examined by immunofluorescent staining for markers of stemness (p63α, ABCG2, CK19 and Vimentin in case of LESCs), proliferation (Ki-67), limbal epithelial- (CK 8/18) and differentiated cornea epithelial- cells (CK3 and 12), and collagen I, IV and V.
Morphological and immunostaining analysis revealed that both cells types when cultured long-term can form stratified 3D tissue layers with a distinct extracellular matrix deposition and organization. The LESCs showed robust expression of p63α, ABCG2 and Vim, low level expression of CK3/12 and a subpopulation of proliferative cells - a clear advantage for their use in future transplantation. Both the LESCs and the CSSCs produced distinct and organized collagen structure.
Overall, we provide a model for generating highly pluripotent, long-standing 3D cultures from LESCs and CSSCs which can be used for further research purposes or transplantation
This PDF is available to Subscribers Only