title: A simple method for establishing adherent ex vivo explant cultures from human eye pathologies for use in subsequent calcium imaging and inflammatory studies.
abstract: A novel, simple, and reproducible method for cultivating pathological tissues obtained from human eyes during surgery was developed using viscoelastic material as a tissue adherent to facilitate cell attachment and expansion and calcium imaging of cultured cells challenged by mechanical and acetylcholine (ACh) stimulation as well as inflammatory studies. Anterior lens capsule-lens epithelial cells (aLC-LECs) from cataract surgery and proliferative diabetic retinopathy (PDR) fibrovascular epiretinal membranes (fvERMs) from human eyes were used in the study. We hereby show calcium signaling in aLC-LECs by mechanical and acetylcholine (ACh) stimulation and indicate presence of ACh receptors in these cells. Furthermore, an ex vivo study model was established for measuring the inflammatory response in fvERMs and aLC-LECs upon TNFα treatment.
authors.affiliation: Eye Hospital, University Medical Centre, 1000 Ljubljana, Slovenia.
authors.forename: Sofija
authors.identifier: 0000-0001-8130-728X
authors.initials: S
authors.lastname: Andjelic
authors.affiliation: Eye Hospital, University Medical Centre, 1000 Ljubljana, Slovenia.
authors.forename: Xhevat
authors.identifier: 0000-0002-9865-4111
authors.initials: X
authors.lastname: Lumi
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, University of Szeged, Korányi fasor 10-11, Szeged 6720, Hungary.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Veréb
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, University of Szeged, Korányi fasor 10-11, Szeged 6720, Hungary.
authors.forename: Natasha
authors.identifier: 0000-0002-3898-9729
authors.initials: N
authors.lastname: Josifovska
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, University of Szeged, Korányi fasor 10-11, Szeged 6720, Hungary.
authors.forename: Andrea
authors.initials: A
authors.lastname: Facskó
authors.affiliation: Eye Hospital, University Medical Centre, 1000 Ljubljana, Slovenia.
authors.forename: Marko
authors.initials: M
authors.lastname: Hawlina
authors.affiliation: Department of Ophthalmology, Faculty of Medicine, University of Szeged, Korányi fasor 10-11, Szeged 6720, Hungary ; Stem Cells and Eye Research Laboratory, Department of Biochemistry and Molecular Biology, Apoptosis and Genomics Research Group of the Hungarian Academy of Sciences, University of Debrecen, Szeged 6720, Hungary.
authors.forename: Goran
authors.identifier: 0000-0003-2905-9252
authors.initials: G
authors.lastname: Petrovski
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D057866:Anterior Capsule of the Lens / Q000473:pathology; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D002386:Cataract / Q000473:pathology; D002387:Cataract Extraction; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism*; D003930:Diabetic Retinopathy / Q000473:pathology; D019773:Epiretinal Membrane / Q000473:pathology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D007908:Lens, Crystalline / Q000473:pathology; D008856:Microscopy, Fluorescence; D046509:Tissue Culture Techniques / Q000295:instrumentation / Q000379:methods*; D055682:Viscoelastic Substances / Q000378:metabolism / Q000494:pharmacology; D018630:Vitreoretinopathy, Proliferative / Q000473:pathology
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators; D055682:Viscoelastic Substances; D000109:Acetylcholine; D002118:Calcium