Highly efficient ex vivo gene delivery into human corneal endothelial cells by recombinant adeno-associated virus.

Purpose: Gene delivery at high efficiency is crucial for cornea endothelial cell gene therapy. This study investigated the efficiency of gene transfer by recombinant adeno-associated virus (rAAV) in an organ culture system.Methods: Human cornea tissue was exposed to rAAV delivering green fluorescent protein (ss-rAAV2-CMV-GFP) for one hour and then cultured at 31ˆC for 2 weeks in a medium supplemented with growth factors. Endothelial cells expressing GFP gene were then identified.Results: High-efficiency gene transfer was found in over 90% of endothelial cells. Gene expression could be detected within 24 hours and remained stable up to 2 weeks in the organ culture system.Conclusions: The high-delivery efficiency and rapid induction of gene expression indicate that rAAV is a promising vector for cornea endothelial cell gene therapy for ocular diseases. Organ culture at 31ˆC using culture medium supplemented with growth factors significantly facilitates gene transfer into human corneal endothelium.