title: Müller and macrophage-like cell interactions in an organotypic culture of porcine neuroretina.
abstract: OBJECTIVE
  To analyze the in vitro Müller cell modifications in an organotypic culture of porcine neuroretina in response to the addition of a blood-derived mononuclear fraction (MNF; monocytes and lymphocytes) as a source of macrophages.
  
  
  METHODS
  Control and MNF-stimulated neuroretinal explants were examined at 3, 6, and 9 days of culture. Specimens were processed for epoxy-resin embedding and cryosectioning. Light and immunofluorescence microscopy were performed, using toluidine blue staining and antibodies against glial fibrillary acidic protein (GFAP), as a reactive gliosis marker, and cellular retinaldehyde-binding protein (CRALBP), as a Müller cell marker.
  
  
  RESULTS
  Compared to controls, explants cocultured with MNF displayed increased cellular disorganization and larger tissue invasion of the subretinal space at 9 days of culture. Immunostaining of the MNF-treated explants revealed evidence of more reactive gliosis and greater number of GFAP-immunoreactive Müller cells that had increased width and processes extending into the subretinal space and forming a multilayer tissue. Astrocytes also responded to the MNF addition, producing extensions that invaded the neuroretinal outer layers.
  
  
  CONCLUSIONS
  Addition of MNF stimulates modifications of Müller cells, producing a wider intraretinal reactive gliosis and tissue proliferation at the subretinal space (outer layers of the retina). These findings emphasize the role of macrophage-like cells in the production of changes in retinal structure observed after retinal detachment in humans.
authors.affiliation: University of Valladolid, Instituto Universitario de Oftalmobiologia Aplicada (IOBA), Campus Miguel Delibes s/n Valladolid, Valladolid, Spain. ifernandezb@ioba.med.uva.es
authors.forename: Ivan
authors.initials: I
authors.lastname: Fernandez-Bueno
authors.forename: Jose Carlos
authors.initials: JC
authors.lastname: Pastor
authors.forename: Manuel Jose
authors.initials: MJ
authors.lastname: Gayoso
authors.forename: Ignacio
authors.initials: I
authors.lastname: Alcalde
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Garcia
mesh_terms: D000818:Animals; D002450:Cell Communication; D018920:Coculture Techniques; D016616:Cryoultramicrotomy; D005455:Fluorescent Antibody Technique; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008264:Macrophages / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D009924:Organ Culture Techniques; D012160:Retina / Q000166:cytology*; D034421:Sus scrofa; D014048:Tolonium Chloride
chemical_list: D014048:Tolonium Chloride