title: Human organotypic retinal cultures (HORCs) as a chronic experimental model for investigation of retinal ganglion cell degeneration.
abstract: There is a growing need for models of human diseases that utilise native, donated human tissue in order to model disease processes and develop novel therapeutic strategies. In this paper we assessed the suitability of adult human retinal explants as a potential model of chronic retinal ganglion cell (RGC) degeneration. Our results confirmed that RGC markers commonly used in rodent studies (NeuN, βIII Tubulin and Thy-1) were appropriate for labelling human RGCs and followed the expected differential expression patterns across, as well as throughout, the macular and para-macular regions of the retina. Furthermore, we showed that neither donor age nor post-mortem time (within 24 h) significantly affected the initial expression levels of RGC markers. In addition, the feasibility of using human post mortem donor tissue as a long-term model of RGC degeneration was determined with RGC protein being detectable up to 4 weeks in culture with an associated decline in RGC mRNA and significant, progressive, apoptotic labelling of NeuN(+) cells. Differences in RGC apoptosis might have been influenced by medium compositions indicating that media constituents could play a role in supporting axotomised RGCs. We propose that using ex vivo human explants may prove to be a useful model for testing the effectiveness of neuroprotective strategies.
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich, NR4 7TJ, UK; John van Geest Centre for Brain Repair, University of Cambridge, Cambridge, CB2 0PY, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Osborne
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich, NR4 7TJ, UK; Department of Ophthalmology, Norfolk and Norwich University Hospital, Norwich, NR4 7UY, UK.
authors.forename: Marina
authors.initials: M
authors.lastname: Hopes
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich, NR4 7TJ, UK.
authors.forename: Phillip
authors.initials: P
authors.lastname: Wright
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich, NR4 7TJ, UK; Department of Ophthalmology, Norfolk and Norwich University Hospital, Norwich, NR4 7UY, UK; School of Biological Sciences, University of East Anglia, Norwich, NR4 7TJ, UK.
authors.forename: David C
authors.initials: DC
authors.lastname: Broadway
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich, NR4 7TJ, UK. Electronic address: j.sanderson@uea.ac.uk.
authors.forename: Julie
authors.initials: J
authors.lastname: Sanderson
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D034961:Antigens, Nuclear / Q000235:genetics; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002470:Cell Survival; D002478:Cells, Cultured; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D020287:In Situ Nick-End Labeling; D008875:Middle Aged; D008962:Models, Theoretical; D009410:Nerve Degeneration / Q000378:metabolism / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012165:Retinal Ganglion Cells / Q000378:metabolism / Q000473:pathology*; D018800:Thy-1 Antigens / Q000235:genetics; D014019:Tissue Donors; D014404:Tubulin / Q000235:genetics
chemical_list: D034961:Antigens, Nuclear; D015415:Biomarkers; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C498585:TUBB3 protein, human; D018800:Thy-1 Antigens; D014404:Tubulin; C506808:neuronal nuclear antigen NeuN, human
keywords: Apoptosis; Degeneration; Glaucoma; Human; Model; RGC; Retina