title: Functional and molecular characterization of ex vivo cultured epiretinal membrane cells from human proliferative diabetic retinopathy.
abstract: Characterization of the cell surface marker phenotype of ex vivo cultured cells growing out of human fibrovascular epiretinal membranes (fvERMs) from proliferative diabetic retinopathy (PDR) can give insight into their function in immunity, angiogenesis, and retinal detachment. FvERMs from uneventful vitrectomies due to PDR were cultured adherently ex vivo. Surface marker analysis, release of immunity- and angiogenesis-pathway-related factors upon TNF α activation and measurement of the intracellular calcium dynamics upon mechano-stimulation using fluorescent dye Fura-2 were all performed. FvERMs formed proliferating cell monolayers when cultured ex vivo, which were negative for endothelial cell markers (CD31, VEGFR2), partially positive for hematopoietic- (CD34, CD47) and mesenchymal stem cell markers (CD73, CD90/Thy-1, and PDGFR β ), and negative for CD105. CD146/MCAM and CD166/ALCAM, previously unreported in cells from fvERMs, were also expressed. Secretion of 11 angiogenesis-related factors (DPPIV/CD26, EG-VEGF/PK1, ET-1, IGFBP-2 and 3, IL-8/CXCL8, MCP-1/CCL2, MMP-9, PTX3/TSG-14, Serpin E1/PAI-1, Serpin F1/PEDF, TIMP-1, and TSP-1) were detected upon TNF α activation of fvERM cells. Mechano-stimulation of these cells induced intracellular calcium propagation representing functional viability and role of these cells in tractional retinal detachment, thus serving as a model for studying tractional forces present in fvERMs in PDR ex vivo.
authors.affiliation: Stem Cells and Eye Research Laboratory, Department of Biochemistry and Molecular Biology, Medical and Health Science Center, Faculty of Medicine, University of Debrecen, Debrecen H-4010, Hungary.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Veréb
authors.forename: Xhevat
authors.initials: X
authors.lastname: Lumi
authors.forename: Sofija
authors.initials: S
authors.lastname: Andjelic
authors.forename: Mojca
authors.initials: M
authors.lastname: Globocnik-Petrovic
authors.forename: Mojca
authors.initials: M
authors.lastname: Urbancic
authors.forename: Marko
authors.initials: M
authors.lastname: Hawlina
authors.forename: Andrea
authors.initials: A
authors.lastname: Facskó
authors.forename: Goran
authors.initials: G
authors.lastname: Petrovski
mesh_terms: D015415:Biomarkers / Q000032:analysis* / Q000378:metabolism; D018929:Cell Culture Techniques; D002470:Cell Survival; D003930:Diabetic Retinopathy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019773:Epiretinal Membrane / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012160:Retina / Q000378:metabolism / Q000473:pathology
chemical_list: D015415:Biomarkers; D008565:Membrane Proteins