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[Gene transfer into corneal endothelial cells by Helios gene gun]
Wen Nan Lu1, Nobuyuki Ebihara, Keiko Fujiki
1Department of Ophthalmology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
Purpose:
To examine the efficiency of particle-mediated gene transfer into rabbit corneal endothelium with a Helios gene gun system.
Methods:
Using gene gun, plasmid DNA coding for green fluorescent protein(GFP) was transferred into cultured rabbit endothelial cells and rabbit corneal grafts from their endothelial site. Then the transferred corneas were transplanted as autografts. The GFP expression was detected by fluorescence microscopy. Histological examination of corneas was performed by light microscope and scanning electron microscope.
Results:
In cultured rabbit endothelial cells, the transfection efficiency without damage to cells was about 7% under optimized helium pressure of 120 psi. In ex vivo, the GFP expression was limited to endothelial cells of the grafts at 140 psi. The histological findings from light and scanning electron microscopy of the grafts showed no severe mechanical damage in endothelial cells penetrated by gold particles.
Conclusions:
The helios gene gun system successfully transferred genes into corneal endothelial cells. However, further studies will be required to obtain the stable results of the particle mediated gene transfer into the endothelial cells of corneal grafts for clinical practice.