Highly efficient ex vivo gene delivery into human corneal endothelial cells by recombinant adeno-associated virus.
OBJECTIVE
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.
Lai
Li
L
Graduate Institute of Clinical Medical Sciences,Chang Gang University, Tau-Yuan, Taiwan.
Lin
Ken
K
Foulks
Gary
G
Ma
Lih
L
Xiao
Xiao
X
Chen
Ko
K
D003673:Defective Viruses; D000229:Dependovirus / Q000235:genetics*; D004728:Endothelium, Corneal / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009924:Organ Culture Techniques; D014162:Transfection / Q000379:methods*
D049452:Green Fluorescent Proteins