title: A novel coculture model of porcine central neuroretina explants and retinal pigment epithelium cells.
abstract: OBJECTIVE
  To develop and standardize a novel organ culture model using porcine central neuroretina explants and RPE cells separated by a cell culture membrane.
  
  
  METHODS
  RPE cells were isolated from porcine eyes, expanded, and seeded on the bottom of cell culture inserts. Neuroretina explants were obtained from the area centralis and cultured alone (controls) on cell culture membranes or supplemented with RPE cells in the same wells but physically separated by the culture membrane. Finally, cellular and tissue specimens were processed for phase contrast, cyto-/histological, and immunochemical evaluation. Neuroretina thickness was also determined.
  
  
  RESULTS
  Compared to the neuroretinas cultured alone, the neuroretinas cocultured with RPE cells maintained better tissue structure and cellular organization, displayed better preservation of photoreceptors containing rhodopsin, lower levels of glial fibrillary acidic protein immunoexpression, and preservation of cellular retinaldehyde binding protein both markers of reactive gliosis. Neuroretina thickness was significantly greater in the cocultures.
  
  
  CONCLUSIONS
  A coculture model of central porcine neuroretina and RPE cells was successfully developed and standardized. This model mimics a subretinal space and will be useful in studying interactions between the RPE and the neuroretina and to preclinically test potential therapies.
authors.affiliation: Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, Valladolid, Spain; Department of Ophthalmology, Hospital Clinico Universitario, Valladolid, Spain.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Di Lauro
authors.affiliation: Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, Valladolid, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Rodriguez-Crespo
authors.affiliation: Department of Cellular Biology, Histology and Pharmacology, University of Valladolid, Valladolid, Spain.
authors.forename: Manuel J
authors.initials: MJ
authors.lastname: Gayoso
authors.affiliation: Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, Valladolid, Spain.
authors.forename: Maria T
authors.initials: MT
authors.lastname: Garcia-Gutierrez
authors.affiliation: Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, Valladolid, Spain; Department of Ophthalmology, Hospital Clinico Universitario, Valladolid, Spain.
authors.forename: J Carlos
authors.initials: JC
authors.lastname: Pastor
authors.affiliation: Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, Valladolid, Spain; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y Leon, Valladolid, Spain.
authors.forename: Girish K
authors.initials: GK
authors.lastname: Srivastava
authors.affiliation: Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, Valladolid, Spain; Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y Leon, Valladolid, Spain.
authors.forename: Ivan
authors.initials: I
authors.lastname: Fernandez-Bueno
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018920:Coculture Techniques; D007150:Immunohistochemistry; D008954:Models, Biological; D009924:Organ Culture Techniques; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism; D013552:Swine
chemical_list: D015415:Biomarkers