title: A Porcine Anterior Segment Perfusion and Transduction Model With Direct Visualization of the Trabecular Meshwork.
abstract: OBJECTIVE
  To establish a consistent and affordable, high quality porcine anterior segment perfusion and transduction model that allows direct visualization of the trabecular meshwork.
  
  
  METHODS
  Porcine anterior segments were cultured within 2 hours of death by removing lens and uvea and securing in a specially designed petri dish with a thin bottom to allow direct visualization of the trabecular meshwork with minimal distortion. Twenty-two control eyes (CO) with a constant flow rate were compared to eight gravity perfused eyes (COgr, 15 mm Hg). We established gene delivery to the TM using eGFP expressing feline immunodeficiency virus (FIV) vector GINSIN at 108 transducing units (TU) per eye (GINSIN_8, n = 8) and 107 TU (GINSIN_7, n = 8). Expression was assessed for 14 days before histology was obtained.
  
  
  RESULTS
  Pig eyes were a reliable source for consistent and high quality anterior segment cultures with a low failure rate of 12%. Control eyes had an intraocular pressure (IOP) of 15.8 ± 1.9 mm Hg at fixed pump perfusion with 3 μL/min compared to gravity perfused COgr with imputed 3.7 ± 1.6 μL/min. Vector GINSIN_8 eyes experienced a transient posttransduction IOP increase of 44% that resolved at 48 hours; this was not observed in GINSIN_7 eyes. Expression was higher in GINSIN_8 than in GINSIN_7 eyes. Trabecular meshwork architecture was well preserved.
  
  
  CONCLUSIONS
  Compared with previously used human donor eyes, this inexpensive porcine anterior segment perfusion model is of sufficient, repeatable high quality to develop strategies of TM bioengineering. Trabecular meshwork could be observed directly. Despite significant anatomic differences, effects of transduction replicate the main aspects of previously explored human, feline and rodent models.
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Ralitsa T
authors.initials: RT
authors.lastname: Loewen
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Pritha
authors.initials: P
authors.lastname: Roy
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Park
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Adrianna
authors.initials: A
authors.lastname: Jensen
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Gordon
authors.initials: G
authors.lastname: Scott
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Devora
authors.initials: D
authors.lastname: Cohen-Karni
authors.affiliation: Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota, United States.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Fautsch
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Joel S
authors.initials: JS
authors.lastname: Schuman
authors.affiliation: Department of Ophthalmology University of Pittsburgh, Pittsburgh, Pennsylvania, United States.
authors.forename: Nils A
authors.initials: NA
authors.lastname: Loewen
mesh_terms: D000818:Animals; D000869:Anterior Eye Segment / Q000378:metabolism* / Q000473:pathology; D001082:Aqueous Humor / Q000378:metabolism*; D002415:Cats; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005901:Glaucoma / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D006801:Humans; D007429:Intraocular Pressure; D010477:Perfusion / Q000379:methods*; D013552:Swine; D014129:Trabecular Meshwork / Q000473:pathology*