The scientists here previously demonstrated the potential of utilizing non-cultured primary corneal endothelial cells isolated from donor corneas with low endothelial cell density for simple non-cultured endothelial cell injection therapy. This study aimed to develop a robust and semi-automated approach for cell counting, characterize the extent of cellular manipulation, and evaluate the translational workflow. To address this, the authors evaluated manual and automated cell counting approaches and characterized the extent of manipulation of CEnCs through the analysis of cell cycle status, gene expressions, and transcriptomic profiles with single-cell RNA-sequencing. Analysis of cell cycle status, cell cycle genes, and transcriptomic profiles revealed close resemblance between the native corneal endothelium and its donor-matched SNEC-I-harvested cells. In conclusion, SNEC-I therapy serves as an attractive corneal endothelial therapeutic approach in view of the minimal extent of cellular manipulation.
