GENE-TRANSFER TO EX-VIVO STORED CORNEAS

Purpose, We examined the efficiency and kinetics of recombinant adenov irus vector-mediated gene transfer to rat and rabbit cornea in culture ex vivo. Methods, A recombinant replication-defective adenovirus was used to transfer a lacZ marker gene to whole rat and rabbit corneas in culture. Histochemistry was used to localise transgene expression and a colorimetric assay to quantify recombinant protein expression. Resu lts. After infection with recombinant virus and culture for 3 days, hi gh-efficiency gene transfer was found, with expression in most endothe lial cells of both species. Minimal expression was found in other corn eal cell types. On histochemistry, longer duration of expression was f ound in rat than in rabbit endothelium. In both rat and rabbit cornea, highest levels of recombinant protein were found at days 3-7 after in cubation with virus, decreasing to low or undetectable levels at 21 da ys. Conclusion, Adenovirus vectors allow high-efficiency transgene exp ression in cornea, largely restricted to the endothelial cells of ex v ivo cultured cornea. Kinetics of expression differ according to the sp ecies of cornea studied, a factor that must be considered if this vect or is used in further studies.