Ex vivo porcine corneal storage using an innovative bioreactor
Abstract
Purpose
There is no animal model of medium-long-term corneal storage for easily available animals because, contrary to human, ex vivo, animal corneas rapidly and dramatically swell and loose their transparency. As intraocular pressure and optimal function of endothelial cells are critically important for corneal homeostasis, we suppose that the passive eye banking technique is not adapted for corneas of young animals with high stromal swelling pressure. We therefore reproduce physiological parameters to improve storage of animal corneas.
Methods
We designed a bioreactor (BR) that restores a pressure equivalent to the intra ocular pressure in the endothelial chamber while allowing continuous renewing of media in both epithelial and endothelial chambers. Epithelial side underwent alternating air-lifting and medium immersion to reproduce blinking. Porcine eyeballs were obtained from a local slaughterhouse within 4 hours after death. Corneas were stored either in a BR or in conventional vials, both in standard organ culture medium. Transparency, thickness, histological structure and immunohistological staining (ABCB5, PAX6, 5-ethynyl-2′-deoxyuridine, K3-K12, Laminin-5) were compared to conventionally stored corneas after 2 weeks.
Results
Porcine corneas stored in bioreactor were more transparent and thinner. Increased endothelial cell viability was observed in the BR and epithelial layer was preserved and mature. Epithelial stem cells also survived.
Conclusions
The porcine version of our BR mimics physiological condition and improves corneal storage. It could be a new model of eye banking and a powerful experimental platform to study corneal physiopathology. Grant: UJM, ANSM.
