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

• Published in: Scientific reports Vol. 10; no. 1; p. 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: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 04.05.2020; Nature Publishing Group
• 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
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-64311-x

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.