Cultured human corneal endothelial cells on a collagen vitrigel carrier as a donor tissue for endothelial keratoplasty

Abstract

Purpose : The goal of this study was to develop a method to culture corneal endothelial cells onto a transparent collagen vitrigel as a donor tissue for EK.

Methods : Primary human corneal endothelial cells (HCECs) were isolated and expanded in vitro using an animal-free medium. A transparent, thin collagen vitrigel was prepared using a three-step process composed of gelation, vitrification, and rehydration, and the collagen vitrigel was used as a carrier to deliver the cultured HCECs. The HCECs on a 7.5 mm collagen vitrigel was brought into the cat anterior chamber after removing the native endothelium and attached to the posterior stroma (HCEC group). Cat corneas with collagen vitrigel transplantation were used as controls (control group).

Results : Propagated HCECs showed expression of markers indicative of the human corneal endothelium. HCECs cultured on collagen vitrigel exhibited cobblestone-like morphology along with similar/enhanced expression of corneal endothelial markers ZO-1 and Na⁺/K⁺-ATPase, compared to identical cultures on plastic tissue culture plate. The cornea was clear in the HCEC group on day 28 after surgery and it was thinning over the 28-day time frame.

Conclusions : This study showed that cell-based therapy for corneal endothelial dysfunction delivered by a collagen vitrigel carrier could be an alternative to traditional corneal transplantation.

This is a 2020 ARVO Annual Meeting abstract.