Abstract
The corneal endothelium maintains corneal transparency by its pump and barrier functions; consequently, its decompensation due to any pathological reason causes severe vision loss due to corneal haziness. Corneal transplantation is the only therapeutic choice for treating corneal endothelial dysfunction, but associated problems, such as a shortages of donor corneas, the difficulty of the surgical procedure, and graft failure, still need to be resolved. Regenerative medicine is attractive to researchers as a means of providing innovative therapies for corneal endothelial dysfunction, as it now does for other diseases. We previously demonstrated the successful regeneration of corneal endothelium in animal models by injecting cultured corneal endothelial cells (CECs) in combination with a Rho kinase (ROCK) inhibitor. The purpose of the present study was to optimize the vehicle for clinical use in cell-based therapy. Our screening of cell culture media revealed that RELAR medium promoted CEC adhesion. We then modified RELAR medium by removing hormones, growth factors, and potentially toxic materials to generate a cell therapy vehicle (CTV) composed of amino acid, salts, glucose, and vitamins. Injection of CECs in CTV enabled efficient engraftment and regeneration of the corneal endothelium in the rabbit corneal endothelial dysfunction model, with restoration of a transparent cornea. The CECs retained >85% viability after a 24 hour preservation as a cell suspension in CTV at 4°C and maintained their potency to regenerate the corneal endothelium in vivo. The vehicle developed here is clinically applicable for cell-based therapy aimed at treating the corneal endothelium. Our strategy involves the generation of vehicle from a culture medium appropriate for a given cell type by removing materials that are not favorable for clinical use.
Highlights
The cornea serves as the window of the eye, and its transparency is critical for vision
We evaluated the feasibility of injecting Corneal endothelial cells (CECs) in cell therapy vehicle (CTV) into a rabbit corneal endothelial dysfunction model, and we showed that the corneal endothelium was regenerated without any adverse effects
We screened the effect of various cell culture media on human CEC (HCEC) adhesion compared with adhesion in OptiMEM-I, a standard culture medium used for HCEC culture by several researchers, Fig 1
Summary
The cornea serves as the window of the eye, and its transparency is critical for vision. One function of the corneal endothelium is the maintenance of corneal transparency, which is PLOS ONE | DOI:10.1371/journal.pone.0158427. Generation of a New Vehicle for Cell-Based Therapy for Treating Corneal Endothelial Dysfunction controlled by the regulation of aqueous humor flow to the corneal stroma by the pump and barrier functions of the corneal endothelium. Corneal endothelial cells (CECs) have very limited proliferative ability and rarely show mitosis in humans after birth [1,2,3]. Severe damage to the corneal endothelium induces irreversible decompensation of endothelial function and leads to corneal haziness. Fuchs endothelial corneal dystrophies and decompensation following cataract surgery are the leading causes of corneal endothelial dysfunction [5]
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