The impact of umbilical cord blood platelet lysate on human corneal endothelium during organ culture.

PURPOSE: To evaluate whether umbilical cord blood platelet lysate (UCB-PL) can preserve the morphology and characteristics of human corneal endothelial cells during organ culture and serve as a xeno-free alternative to fetal bovine serum (FBS).

METHODS

Paired human donor corneas were cultured for 28 days in α-MEM-based medium supplemented with either 2% FBS or 2% UCB-PL. Endothelial morphology, cell density, viability, and mosaic regularity were assessed at predefined time points. Metabolic activity was evaluated by measuring pH, glucose, and lactate concentrations in the culture media. Endothelial integrity, apoptosis, and proliferation were analyzed by whole-mount immunofluorescence staining for ZO-1, Na⁺/K⁺-ATPase, caspase-3, and Ki-67.

RESULTS

Corneas stored in UCB-PL-supplemented medium demonstrated endothelial morphology, cell density decline, and viability comparable to those observed in FBS-supplemented medium throughout the culture period. No significant differences were detected between groups at any time point. Metabolic analysis showed sustained glucose availability and expected lactate accumulation in both media, without evidence of nutrient depletion. Immunofluorescence confirmed preserved endothelial junctional organization and pump protein expression, with no signs of endothelial apoptosis or proliferation.

CONCLUSION

In this pilot feasibility study, UCB-PL supported preservation of human corneal endothelial morphology and endothelial-associated protein expression during organ culture and demonstrated similar trends to FBS. These findings support further investigation of UCB-PL as a potential xeno-free alternative for corneal storage media.