July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Müller glia isolated from human iPSC/ESC derived retinal organoids produce neuroprotective antioxidants against oxidative stress
Author Affiliations & Notes
  • Weixin Wang
    NIHR Biomedical Research Centre for Ophthalmology, UCL Institute of Ophthalmology and Moorfields Eye Hospital, London, UK, London, ENGLAND, United Kingdom
  • Karen Eastlake
    NIHR Biomedical Research Centre for Ophthalmology, UCL Institute of Ophthalmology and Moorfields Eye Hospital, London, UK, London, ENGLAND, United Kingdom
  • Celia Murray-Dunning
    NIHR Biomedical Research Centre for Ophthalmology, UCL Institute of Ophthalmology and Moorfields Eye Hospital, London, UK, London, ENGLAND, United Kingdom
  • Erika Aquino
    NIHR Biomedical Research Centre for Ophthalmology, UCL Institute of Ophthalmology and Moorfields Eye Hospital, London, UK, London, ENGLAND, United Kingdom
  • Peter J Coffey
    NIHR Biomedical Research Centre for Ophthalmology, UCL Institute of Ophthalmology and Moorfields Eye Hospital, London, UK, London, ENGLAND, United Kingdom
  • G. Astrid Limb
    NIHR Biomedical Research Centre for Ophthalmology, UCL Institute of Ophthalmology and Moorfields Eye Hospital, London, UK, London, ENGLAND, United Kingdom
  • Footnotes
    Commercial Relationships   Weixin Wang, None; Karen Eastlake, None; Celia Murray-Dunning, None; Erika Aquino, None; Peter Coffey, None; G. Astrid Limb, None
  • Footnotes
    Support  MRC and NIHR Biomedical Research Centre at Moorfields Eye Hospital and the UCL Institute of Ophthalmology
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 1495. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Weixin Wang, Karen Eastlake, Celia Murray-Dunning, Erika Aquino, Peter J Coffey, G. Astrid Limb; Müller glia isolated from human iPSC/ESC derived retinal organoids produce neuroprotective antioxidants against oxidative stress. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1495.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : Intravitreal transplantation of Müller cells into animal models of retina degeneration has shown that these cells partially restore visual function without integration into the neural retina. This effect has been ascribed to the neuroprotective effects and metabolic support of Müller glia to retinal neurons. They produce antioxidant molecules to protect neurons against oxidative stress and glutamate toxicity. Because of the need to produce Müller cell populations that can be used therapeutically, we have isolated Müller glia from retinal organoids formed by human iPSC/ESC, examined the antioxidant properties of these cells and compared them with the human Müller cell line MIO-M1.

Methods : Retinal organoids were derived from human ESC line Shef6 and iPSC line BJ using slight modifications of method published by Nakano et al., 2012. Müller glia were isolated by papain dissociation at various time points after initiation of organoid differentiation and propagated in vitro on fibronectin coated plates. The identity of these cells was confirmed by mRNA and protein expression of known Müller glia markers using RT-PCR, Western blotting and immunocytochemistry. Gene expression of various antioxidants produced by Müller glia was examined by RT-PCR. Quantification of antioxidants PRD6, PON2 and HO1 released by cells in culture was performed by ELISA.

Results : The results showed that purified populations of Müller glia could be obtained from human iPSC/ESC derived retina organoids. These cells expressed Müller markers including CRALBP, Nestin, Vimentin, Glutamine synthetase, Sox9, Pax6 and Notch1 as confirmed by RT-PCR, Western blotting and immunostaining. They consistently expressed mRNA encoding the atioxidants PRD6, FOHL1, CQ10A, GST, GSR, LIAS and HO1. SOD2 mRNA was not detected in organoid-derived Müller glia but was observed in MIO-M1 cells. Interestingly, ELISA data showed that organoid-derived Müller glia released high levels of the antioxidants PRD6 and HO1 into the culture supernatant, whilst PON2 was not detected.

Conclusions : Our findings demonstrate that retinal organoid derived Müller glia display potent antioxidant ability as judged by the gene expression of antioxidants and the release of antioxidants into culture supernatant. These cells hold promising potential for in vivo transplantation to repair the neural retina with the neuroprotective antioxidants.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×