July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Novel Retina Specific Laminin Isoforms Recapitulate Retinal Interphotoreceptor Matrix to Generate Human Embryonic Stem Cell-derived Photoreceptors
Author Affiliations & Notes
  • Hwee Goon Tay
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
  • Zuhua Cai
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
  • Yang Sun
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
  • Jing Guo
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
    Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
  • Aida Moreno-Moral
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
  • Wei Sheng Tan
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
  • Karl Tryggvason
    Cardiovascular and Metabolic Disorders, DUKE NUS Medical School, Singapore, Singapore
    Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden
  • Footnotes
    Commercial Relationships   Hwee Goon Tay, None; Zuhua Cai, None; Yang Sun, None; Jing Guo, None; Aida Moreno-Moral, None; Wei Sheng Tan, None; Karl Tryggvason, None
  • Footnotes
    Support  Young Investigator Research Grant (YIRG) - National Medical Research Council (NMRC) of Singapore
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 61. doi:
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      Hwee Goon Tay, Zuhua Cai, Yang Sun, Jing Guo, Aida Moreno-Moral, Wei Sheng Tan, Karl Tryggvason; Novel Retina Specific Laminin Isoforms Recapitulate Retinal Interphotoreceptor Matrix to Generate Human Embryonic Stem Cell-derived Photoreceptors. Invest. Ophthalmol. Vis. Sci. 2018;59(9):61.

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

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Abstract

Purpose : Photoreceptors could play a crucial role in retinal cell therapy to treat vision loss. The use of retinal organoid has been hailed to be the established in vitro retinal differentiation method to generate photoreceptors from the pluripotent stem cells. However, this approach is evidently limited by its reproducibility and efficiency level during retinal organoidogenesis. Furthermore, the existing retinal organoid based protocols are not chemically defined and xenogen-free, which include the use of non-human animal serum and Matrigel®, a murine Engelbreth-Holm-Swarm sarcoma extract that are not considered clinically safe. Therefore, it is important to generate clinically safe functional photoreceptors that do not compromise patient’s vision conditions.

Methods : Currently, we have developed a retinal organoid free method to robustly generate photoreceptor progenitors from human embryonic stem cells (hESCs). Our novel alternative approach employs the use of the human recombinant retina-specific laminin isoforms to recapitulate the retinal interphotoreceptor matrix environment. With the support of an analogous retina matrix like surface, the hESCs are being efficiently differentiated towards photoreceptor progenitor-like cells. Unlike retinal organoid, this chemically defined and xenogen-free method does not involve re-plating and manual dissections.

Results : This laminin based differentiation method consistently generates photoreceptor progenitors in every single batch of differentiation. These hESCs-derived photoreceptors are shown to be positive for PAX6, VSX2, CRX and RCVRN as early as Day 30, based on transcriptome (n = 9, p<0.001) and immunocytochemical analyses (n=9). In contrast, the pluripotency and teratoma transcript markers are shown to be drastically downregulated, suggesting reduced associated risk of teratoma formation.

Conclusions : We provide an alternative retina laminin based differentiation method that does not require the formation of retinal organoid. This xenogen-free and chemically defined protocol is compatible with GMP (Good Manufacturing Practice) condition and reproducibly promotes hESCs to photoreceptor progenitor lineage. Hence, these results suggest that our method may constitute an important step towards the future use of hESC-derived photoreceptors to treat vision loss.

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

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