Phenotypic and functional characterization of corneal endothelial cells during in vitro expansion

The advent of cell culture-based methods for the establishment and expansion of human corneal endothelial cells (CEnC) has provided a source of transplantable corneal endothelium, with a significant potential to challenge the one donor-one recipient paradigm. However, concerns over cell identity rem...

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Bibliographic Details
Published in:Scientific reports 2020-05, Vol.10 (1), p.7402-7402, Article 7402
Main Authors: Frausto, Ricardo F., Swamy, Vinay S., Peh, Gary S. L., Boere, Payton M., Hanser, E. Maryam, Chung, Doug. D., George, Benjamin L., Morselli, Marco, Kao, Liyo, Azimov, Rustam, Wu, Jessica, Pellegrini, Matteo, Kurtz, Ira, Mehta, Jodhbir S., Aldave, Anthony J.
Format: Article
Language:English
Subjects:
631/114
631/337/2019
631/80
631/80/509
631/80/79
Adolescent
Adult
BASIC BIOLOGICAL SCIENCES
Cell adhesion
Cell biology
Cell culture
Cell Culture Techniques - methods
Cell Movement
Cell Proliferation
Cellular Senescence
Child
Child, Preschool
Computational Biology
Computational biology and bioinformatics
Cornea
Corneal Transplantation
Endothelial cells
Endothelium
Endothelium, Corneal - cytology
Female
Humanities and Social Sciences
Humans
Male
multidisciplinary
Phenotype
Phenotypes
Science
Science & Technology - Other Topics
Science (multidisciplinary)
Senescence
Signal Transduction
Transcriptome
Transcriptomics
Young Adult
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Summary:The advent of cell culture-based methods for the establishment and expansion of human corneal endothelial cells (CEnC) has provided a source of transplantable corneal endothelium, with a significant potential to challenge the one donor-one recipient paradigm. However, concerns over cell identity remain, and a comprehensive characterization of the cultured CEnC across serial passages has not been performed. To this end, we compared two established CEnC culture methods by assessing the transcriptomic changes that occur during in vitro expansion. In confluent monolayers, low mitogenic culture conditions preserved corneal endothelial cell state identity better than culture in high mitogenic conditions. Expansion by continuous passaging induced replicative cell senescence. Transcriptomic analysis of the senescent phenotype identified a cell senescence signature distinct for CEnC. We identified activation of both classic and new cell signaling pathways that may be targeted to prevent senescence, a significant barrier to realizing the potential clinical utility of in vitro expansion.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-64311-x