Multimodal imaging quality control of epithelia regenerated with cultured human donor corneal limbal epithelial stem cells

Current imaging techniques for the characterization of differentiated corneal limbal stem cells are destructive and cannot be used in eye bank for monitoring the regenerated epithelium in culture. We presented a minimally invasive, multimodal, marker-free imaging method for the investigation of epit...

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Bibliographic Details
Published in:Scientific reports 2017-07, Vol.7 (1), p.5154-10, Article 5154
Main Authors: Lombardo, Marco, Serrao, Sebastiano, Barbaro, Vanessa, Di Iorio, Enzo, Lombardo, Giuseppe
Format: Article
Language:English
Subjects:
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692/308/2171
692/700/1421/2025
Biomaterials
Cell culture
Cell Culture Techniques - methods
Cells, Cultured
Cornea
Epithelial cells
Epithelium
Epithelium, Corneal - cytology
Eye
Humanities and Social Sciences
Humans
Limbus Corneae - cytology
Microscopy, Fluorescence, Multiphoton
multidisciplinary
Multimodal Imaging - methods
Packing
Quality control
Regeneration
Science
Science (multidisciplinary)
Stem cell transplantation
Stem cells
Tissue Donors
Tissue Scaffolds
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Summary:Current imaging techniques for the characterization of differentiated corneal limbal stem cells are destructive and cannot be used in eye bank for monitoring the regenerated epithelium in culture. We presented a minimally invasive, multimodal, marker-free imaging method for the investigation of epithelia regenerated with cultured human donor corneal limbal epithelial stem cells. Two-photon fluorescence and harmonic generation signals were collected from specimens in culture and used for evaluating the structure and morphology of epithelia cultured on two different bio-scaffolds; in addition, donor human corneal tissues were used as controls. The method provided reliable information on the organization of cellular and extracellular components of biomaterial substrates and was highly sensitive to determine differences between the density packing arrangement of epithelial cells of different biomaterials without relying on inferences from exogenous labels. The present minimally invasive standardized quality control methodology can be reliably translated to eye banks and used for monitoring harvested corneal limbal stem cells growth and differentiation in bioengineered materials.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-017-05486-8