RT info:eu-repo/semantics/article
T1 Human corneal epithelial cells harvested from advanced surface ablation (ASA): An optimized in vitro culture protocol
A1 Gutiérrez, Helen
A1 López García, Antonio
A1 Maldonado López, Miguel José
A1 García Posadas, Laura
K1 Corneal epithelium
K1 Cornea
K1 Ablation surgery
K1 Refractive surgery
K1 Collagen
K1 Primary cultures
K1 Trehalose
K1 3201.09 Oftalmología
AB Purpose: Corneal epithelial cells obtained from advanced surface ablation (ASA) surgery provide a valuableresource for in vitro models of ocular surface diseases. The aim of this study is to enhance culture conditions andcharacterize the functionality of EpiASA cells in culture, focusing on their ability to keep the distinctive prop-erties of corneal epithelial cells.Methods: EpiASA samples from 51 patients were included in the study. Two different collection media weretested, and their effect on sample preservation and initial viability was evaluated. Then, cells were disaggregatedand cultured using different strategies to increase cell viability, which was measured by AlamarBlue assay. Oncethe optimized conditions were established, cells were cultured and passaged, and structural and functionalcharacterization of native tissue, primary cultures, and first-passage cultures was performed using atomic forcemicroscopy (AFM), qPCR, and immunofluorescence stainings.Results: The addition of trehalose to the basal collection medium increased EpiASA initial viability. Culturesurface coating with type I collagen, along with the supplementation of culture medium with hydrocortisone,significantly increased cell viability. On the contrary, co-cultures with different ocular cell lines, or the use ofhuman serum, did not provide a sustained benefit. Further low-concentration trehalose supplementation ofEpiASA cultures enhanced monolayer formation and allowed subculturing. AFM and immunofluorescenceconfirmed that passage 1 EpiASA cells retained corneal epithelial characteristics, including well-organizedmicrovilli and uniform expression of barrier and epithelial markers.Conclusion: This research provides an optimized protocol (EpiKeraMAX) for using EpiASA samples for in vitrostudies of human corneal cells
PB Elsevier
SN 0014-4835
YR 2025
FD 2025
LK https://uvadoc.uva.es/handle/10324/78405
UL https://uvadoc.uva.es/handle/10324/78405
LA eng
NO Experimental Eye Research, 2025, vol. 258, p. 110510
NO Producción Científica
DS UVaDOC
RD 11-oct-2025