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  • Title: Highly efficient ex vivo gene delivery into human corneal endothelial cells by recombinant adeno-associated virus.
    Author: Lai L, Lin K, Foulks G, Ma L, Xiao X, Chen K.
    Journal: Curr Eye Res; 2005 Mar; 30(3):213-9. PubMed ID: 15804747.
    Abstract:
    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 31 degrees C 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 31 degrees C using culture medium supplemented with growth factors significantly facilitates gene transfer into human corneal endothelium.
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