A single-cell RNA-seq analysis unravels the heterogeneity of primary cultured human corneal endothelial cells

• Published in: Scientific reports 2023-06, Vol.13 (1), p.9361-9361, Article 9361
• Main Authors: Català, Pere, Groen, Nathalie, LaPointe, Vanessa L. S., Dickman, Mor M.
• Format: Article
• Language: English
• Subjects: 631/337; 631/80; Cell culture; Cell therapy; Cornea; Endothelial cells; Heterogeneity; Humanities and Social Sciences; multidisciplinary; Opacity; Science; Science (multidisciplinary); Transcriptomics
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-36567-6

The cornea is a transparent and avascular tissue located in front of the eye. Its inner surface is lined by a monolayer of corneal endothelial cells (CECs), which maintain the cornea transparency. CECs remain arrested in a non-proliferative state and damage to these cells can compromise their function leading to corneal opacity. The primary culture of donor-derived CECs is a promising cell therapy. It confers the potential to treat multiple patients from a single donor, alleviating the global donor shortage. Nevertheless, this approach has limitations preventing its adoption, particularly culture protocols allow limited expansion of CECs and there is a lack of clear parameters to identify therapy-grade CECs. To address this limitation, a better understanding of the molecular changes arising from the primary culture of CECs is required. Using single-cell RNA sequencing on primary cultured CECs, we identify their variable transcriptomic fingerprint at the single cell level, provide a pseudo-temporal reconstruction of the changes arising from primary culture, and suggest markers to assess the quality of primary CEC cultures. This research depicts a deep transcriptomic understanding of the cellular heterogeneity arising from the primary expansion of CECs and sets the basis for further improvement of culture protocols and therapies.