[Visualization of keratocytes in the human cornea with fluorescence microscopy]

Area of Science:

Context:

Limited data exists on human keratocyte viability during long-term cornea storage.
Stromal keratocytes are often overlooked in current cornea banking culture conditions.
Corneal transplant success is influenced by the health of donor tissue, including keratocytes.

Purpose:

To investigate the density, morphology, and vitality of human keratocytes in stored corneas.
To compare human keratocyte characteristics with porcine models.
To assess the impact of long-term storage on keratocyte health.

Summary:

Human and porcine corneas were stained using the Live/Dead Assay with calcein AM (living cells, green fluorescence) and ethidium homodimer (dead cells, red fluorescence).
Three keratocyte subpopulations were identified in human corneas, with morphological and staining differences compared to porcine corneas, particularly in central and posterior stroma.
Long-term storage resulted in observed cell degeneration, including loss of cytoplasmic processes and an increased number of dead keratocytes.

Impact:

Findings reveal significant cell degeneration in human keratocytes after prolonged storage.
This research underscores the critical need to optimize culture conditions in cornea banking to preserve keratocyte viability.
Improving keratocyte health in stored corneas may enhance corneal transplant survival rates and surgical success.