title: Development of a murine ocular posterior segment explant culture for the study of intravitreous vector delivery.
abstract: The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: Nora
authors: N
authors: Denk
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: Vikram
authors: V
authors: Misra
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: Lynne S
authors: LS
authors: Sandmeyer
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: Bianca B
authors: BB
authors: Bauer
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: Jaswant
authors: J
authors: Singh
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: George W
authors: GW
authors: Forsyth
authors: Department of Small Animal Clinical Sciences (Denk, Sandmeyer, Bauer, Grahn); Veterinary Microbiology (Misra); and Veterinary Biomedical Sciences (Singh, Forsyth), Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan.
authors: Bruce H
authors: BH
authors: Grahn
mesh_terms: D000818:Animals; D002470:Cell Survival; D015316:Genetic Therapy; D005822:Genetic Vectors; D066298:In Vitro Techniques; D016086:Lentivirus; D051379:Mice; D008810:Mice, Inbred C57BL; D057972:Posterior Eye Segment / Q000502:physiology*; D012160:Retina / Q000502:physiology*; D046509:Tissue Culture Techniques; D014161:Transduction, Genetic