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Titolo:
Gene transfer to ovine corneal endothelium
Autore:
Klebe, S; Sykes, PJ; Coster, DJ; Bloom, DC; Williams, KA;
Indirizzi:
Flinders Univ S Australia, Flinders Med Ctr, Dept Ophthalmol, Bedford Pk, SA 5042, Australia Flinders Univ S Australia Bedford Pk SA Australia 5042 SA 5042, Australia Flinders Univ S Australia, Dept Haematol & Genet Pathol, Bedford Pk, SA 5042, Australia Flinders Univ S Australia Bedford Pk SA Australia 5042 SA 5042, Australia Univ Florida, Coll Med, Dept Mol Genet & Microbiol, Gainesville, FL USA Univ Florida Gainesville FL USA l Genet & Microbiol, Gainesville, FL USA
Titolo Testata:
CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY
fascicolo: 5, volume: 29, anno: 2001,
pagine: 316 - 322
SICI:
1442-6404(200110)29:5<316:GTTOCE>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
HERPES-SIMPLEX VIRUS; GRAFT-REJECTION; NONVIRAL VECTOR; OCULAR-TISSUES; THERAPY; CELLS; ADENOVIRUS; DELIVERY; TRANSPLANTATION; EXPRESSION;
Keywords:
corneal endothelium; gene therapy; non-viral and viral vectors; reporter gene;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Williams, KA Flinders Univ S Australia, Flinders Med Ctr, Dept Ophthalmol,Bedford Pk, SA 5042, Australia Flinders Univ S Australia Bedford Pk SA Australia 5042 ralia
Citazione:
S. Klebe et al., "Gene transfer to ovine corneal endothelium", CLIN EXP OP, 29(5), 2001, pp. 316-322

Abstract

Purpose: Modification of a donor cornea by gene therapy has potential to modulate irreversible rejection, the major cause of corneal graft failure. The sheep is a useful model for the human in this respect, as ovine endothelial cells are amitotic. The aim of the study was to investigate the abilityof various non-viral and viral agents to transfer a reporter gene to ovinecorneal endothelium. Methods: The non-viral agents Transfectin-10, Transfectin-20, Transfectin-50, SuperFect, Effectene and CLONfectin were used to deliver the reporter gene, Escherichia coli lacZ, to ovine corneal endothelium in vitro. A Herpessimplex virus-1 and an adenoviral vector each encoding E. coli lacZ were similarly tested. Infected corneas were organ-cultured for up to 7 days in vitro to allow transfection efficiency, duration of gene expression and toxicity attributable to each vector to be compared. Results: Scattered single or clusters of endothelial cells expressing the reporter gene were observed after transfection with CLONfectin, Transfectin-10, Transfectin-20 and Transfectin-50. SuperFect and Effectene were virtually in-effective. At best, the absolute number of infected cells per endothelial monolayer after 3 or 7 days of organ culture was estimated as < 0.01%. The Herpes simplex virus-1 vector also failed to transduce ovine corneal endothelium efficiently. In contrast, transfection rates of up to 70% of endothelial cells were observed with the adenoviral vector. Conclusion: Non-viral vectors and Herpes simplex virus-1 are unlikely to be suitable for gene therapy of corneal endothelium, because the efficiency of transfection is low compared with the rates achieved with adenoviral vectors.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/03/20 alle ore 20:08:25