title: Human limbal neurospheres prevent photoreceptor cell death in a rat model of retinal degeneration.
abstract: BACKGROUND
  The culture of retinal progenitors from an accessible adult stem cell source such as the limbus could provide a useful autologous source of retinal cell therapies. The human corneoscleral limbus contains multipotent stem cells that can be cultured as floating neurospheres. Previous work in rodents has demonstrated neuronal and photoreceptor differentiation from limbal neurosphere cultures. Here, this study has examined undifferentiated cultured adult human limbal neurospheres as donor cells for retinal cell therapies by transplantation into a rat model of retinal degeneration.
  
  
  METHODS
  Gene expression in limbal neurospheres was examined by immunostaining and western blot. Human limbal neurospheres were transplanted into the subretinal space of Royal College of Surgeon's rats. Rats were monitored by optical coherence tomography for 6 weeks then processed for retinal histology.
  
  
  RESULTS
  Human limbal neurospheres expressed the neural lineage markers, Nestin, sex determining region box-2 and N-cadherin, and the retinal transcription factors microphthalmia-associated transcription factor, sex determining region box-2 and orthodentical homeobox-2. Human limbal neurospheres could be cultured to express NeuN, neurofilament and rhodopsin. Rats receiving saline or no injection underwent complete degeneration of the retinal outer nuclear layer after 3 weeks. In contrast, rats injected with human limbal neurospheres or retinal pigment epithelial cells maintained the outer nuclear layer for up to 6 weeks. Gene expression in transplanted limbal neurospheres was inconsistent with the production of mature retinal pigment epithelial or photoreceptor cells.
  
  
  CONCLUSIONS
  Human limbal neurospheres represent an accessible source of autologous donor cells for the treatment of retinal diseases.
authors.affiliation: Centre for Ophthalmology and Visual Science, The University of Western Australia, Perth, Western Australia, Australia.|Ocular Tissue Engineering Laboratory, Lions Eye Institute, Nedlands, Western Australia, Australia.
authors.forename: Samuel
authors.initials: S
authors.lastname: McLenachan
authors.affiliation: Ocular Tissue Engineering Laboratory, Lions Eye Institute, Nedlands, Western Australia, Australia.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Ocular Tissue Engineering Laboratory, Lions Eye Institute, Nedlands, Western Australia, Australia.|Faculty of Pharmacy, Guangxi University of Chinese Medicine, Nanning, Guangxi, China.
authors.forename: Erwei
authors.initials: E
authors.lastname: Hao
authors.affiliation: Ocular Tissue Engineering Laboratory, Lions Eye Institute, Nedlands, Western Australia, Australia.|Faculty of Pharmacy, Guangxi University of Chinese Medicine, Nanning, Guangxi, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Ocular Tissue Engineering Laboratory, Lions Eye Institute, Nedlands, Western Australia, Australia.
authors.forename: Shang-Chih
authors.initials: SC
authors.lastname: Chen
authors.affiliation: Centre for Ophthalmology and Visual Science, The University of Western Australia, Perth, Western Australia, Australia.|Ocular Tissue Engineering Laboratory, Lions Eye Institute, Nedlands, Western Australia, Australia.|Department of Ophthalmology, Royal Perth Hospital, Perth, Western Australia, Australia.
authors.forename: Fred K
authors.identifier: http://orcid.org/0000-0003-2809-9930
authors.initials: FK
authors.lastname: Chen
mesh_terms: D000818:Animals; D034961:Antigens, Nuclear / Q000096:biosynthesis / Q000235:genetics; D015153:Blotting, Western; D016923:Cell Death; D002454:Cell Differentiation; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D005801:Genes, Homeobox / Q000235:genetics; D006801:Humans; D056965:Injections, Intraocular; D016850:Limbus Corneae / Q000166:cytology* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics; D010857:Pigment Epithelium of Eye / Q000378:metabolism / Q000473:pathology*; D051381:Rats; D017949:Retinal Cone Photoreceptor Cells / Q000378:metabolism / Q000473:pathology*; D012162:Retinal Degeneration / Q000235:genetics / Q000473:pathology* / Q000601:surgery; D012243:Rhodopsin / Q000096:biosynthesis / Q000235:genetics; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D041623:Tomography, Optical Coherence
chemical_list: D034961:Antigens, Nuclear; D009419:Nerve Tissue Proteins; C546339:Rbfox3 protein, rat; D004247:DNA; D012243:Rhodopsin
keywords: Royal College of Surgeon's rat; cell transplantation; inherited retinal dystrophy; limbal stem cell; neurosphere