June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Charactering Y79 Cells as a Model of Human Photoreceptor Differentiation
Author Affiliations & Notes
  • Molly John
    Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Ariel Kantor
    Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Michelle E McClements
    Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Robert E MacLaren
    Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Kanmin Xue
    Nuffield Department of Clinical Neuroscience, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Footnotes
    Commercial Relationships   Molly John None; Ariel Kantor None; Michelle McClements None; Robert MacLaren None; Kanmin Xue None
  • Footnotes
    Support  Wellcome Trust: 216593/Z/19/Z, NIHR Oxford Biomedical Research Centre
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3736 – F0342. doi:
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    • Get Citation

      Molly John, Ariel Kantor, Michelle E McClements, Robert E MacLaren, Kanmin Xue; Charactering Y79 Cells as a Model of Human Photoreceptor Differentiation. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3736 – F0342.

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

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Abstract

Purpose : Model systems for the study of human photoreceptor differentiation are limited. These include retinal organoids and a limited number of cell lines which vary in their states of differentiation. Y79 cells are derived from a retinoblastoma in a 2.5-year-old Caucasian female and grow as multicellular clusters in suspension. Here, we aim to determine the pattern of expression of photoreceptor-specific genes in Y79 cells to determine their suitability as a model for studying human cone versus rod fate determination in vitro.

Methods : Y79 cells were cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum, 1% L-glutamine, 1% Pen-Strep in 5% CO2 at 35C. Cells were harvested at passage number 15 for total RNA extraction. The expression of a panel of cone-specific, rod-specific, and pan-photoreceptor genes were analysed by RT-PCR and RT-qPCR (n=3).

Results : Y79 cells demonstrated consistent (n=3) and clear expression of rod-specific (NR2E3, NRL, PDE6A, GNAT1) and cone-specific (ARR3, GNAT2, GNTG2) genes, as well as the pan-photoreceptor transcription factor, CRX. While rhodopsin expression was absent, cone photopigment expression was limited, with minimal expression of medium- and long-wavelength opsins and no detectable short-wavelength opsin by both RT-PCR and qPCR.

Conclusions : Retinoblastomas are generally thought to arise from post-mitotic cone precursors and that cone-related gene circuitry sensitises the cells to the oncogenic effects of RB1 mutations. Our results show that Y79 cells express M/L-cone-specific markers, but also a range of distinct rod markers, which could indicate their origin from a precursor stage during cone/rod fate divergence or a degree of trans-differentiation during oncogenic transformation. Further studies will evaluate the use of Y79 cells as a model system for studying photoreceptor differentiation.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

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