July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Decellularised extracellular matrix from neural retina and retinal pigment epithelium enhance the expression of synaptic markers and light responsiveness of human pluripotent stem cell derived retinal organoids
Author Affiliations & Notes
  • Majlinda Lako
    Institute of Genetic Medicine, Newcastle University, Newcastle, ENGLAND, United Kingdom
  • Birthe Dorgau
    Institute of Genetic Medicine, Newcastle University, Newcastle, ENGLAND, United Kingdom
  • Majed Felemban
    Institute of Genetic Medicine, Newcastle University, Newcastle, ENGLAND, United Kingdom
  • Gerrit Hilgen
    Institute of Neuroscience, Newcastle University, United Kingdom
  • Martin Kiening
    Institute of Genetic Medicine, Newcastle University, Newcastle, ENGLAND, United Kingdom
  • Dean Hallam
    Institute of Genetic Medicine, Newcastle University, Newcastle, ENGLAND, United Kingdom
  • Nicola Claire Hunt
    Institute of Genetic Medicine, Newcastle University, Newcastle, ENGLAND, United Kingdom
  • Yuchun Ding
    School of Computing, United Kingdom
  • Natalio Krasnogor
    School of Computing, United Kingdom
  • Jumana Al-Aama
    PACER, Saudi Arabia
  • Hani Z Asfour
    PACER, Saudi Arabia
  • Evelyne Sernagor
    Institute of Neuroscience, Newcastle University, United Kingdom
  • Footnotes
    Commercial Relationships   Majlinda Lako, None; Birthe Dorgau, None; Majed Felemban, None; Gerrit Hilgen, None; Martin Kiening, None; Dean Hallam, None; Nicola Hunt, None; Yuchun Ding, None; Natalio Krasnogor, None; Jumana Al-Aama, None; Hani Asfour, None; Evelyne Sernagor, None
  • Footnotes
    Support  European Research Council
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 3322. doi:
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      Majlinda Lako, Birthe Dorgau, Majed Felemban, Gerrit Hilgen, Martin Kiening, Dean Hallam, Nicola Claire Hunt, Yuchun Ding, Natalio Krasnogor, Jumana Al-Aama, Hani Z Asfour, Evelyne Sernagor; Decellularised extracellular matrix from neural retina and retinal pigment epithelium enhance the expression of synaptic markers and light responsiveness of human pluripotent stem cell derived retinal organoids. Invest. Ophthalmol. Vis. Sci. 2019;60(9):3322.

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

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Abstract

Purpose : Tissue specific extracellular matrices (ECM) provide structural support and enable access to molecular signals and metabolites which are essential for directing stem cell renewal and differentiation. To mimic this phenomenon in vitro, tissue decellularisation approaches have been developed, resulting in the generation of natural ECM scaffolds that have comparable physical and biochemical properties of the natural tissues. In this study, RPE conditioned medium (CM RPE), decellularised ECM from neural retina (decel NR) and retinal pigment epithelium (decel RPE) were successfully generated and their impacts on differentiation of human pluripotent stem cells (hPSCs) to retinal organoids was investigated.

Methods : CM RPE was collected every day from polarized mature RPE derived from hPSCs while decel NR and RPE were performed using a protocol published by Medberry et al., 2013 from 4 adult bovine eyes and used as supplements to culture media. Retinal organoids were generated from hPSCs following a protocol described in Mellough et al., 2015 with the addition of ROCK inhibitor (Y-27632 dihydrochloride) for the first 48 hours of differentiation. The impacts of CM RPE, decel NR and decel RPE on the on differentiation process was studied using a number of laboratory techniques including: immunohistochemistry, qRT-PCR, electrophysiological recordings and transmission electron microscopy.

Results : Culture media supplementation with decel RPE enhanced RPE generation. Supplementation with decel RPE and CM RPE significantly increased the generation of rod photoreceptors, whilst addition of decel NR and decel RPE significantly enhanced ribbon synapse marker expression and the light responsiveness of hPSC-derived retinal organoids.

Conclusions : This study indicates that decellularised ECM generated from neural retina and RPE plays an important role in photoreceptor maturation, synaptogenesis and light responsiveness of pluripotent stem cell derived retinal organoids. The decellularised ECMs are easy to generate, amenable to scale up and 3D culture conditions, opening up new possibilities for generation of retinal organoids which mimic adult human retina both in cell type composition and function.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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