July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Enhancing Retinal Ganglion Cell Production from Human Pluripotent Stem Cell-Derived 3D Retinal Cultures
Author Affiliations & Notes
  • Xiangmei Zhang
    Ophthalmology, Jules Stein Eye Institute-UCLA, Los Angeles, California, United States
  • Kevin Nguyen
    Ophthalmology, Jules Stein Eye Institute-UCLA, Los Angeles, California, United States
  • Steven A Barnes
    Physio/Biophys/Ophthal/Vis Sci, UCLA, Los Angeles, California, United States
    Physio/Biophys/Ophthal/Vis Sci, Dalhousie University-DAL-11762, Halifax, Nova Scotia, Canada
  • Xian-Jie Yang
    Ophthalmology, Jules Stein Eye Institute-UCLA, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Xiangmei Zhang, None; Kevin Nguyen, None; Steven Barnes, None; Xian-Jie Yang, None
  • Footnotes
    Support  NIH
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 564. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Xiangmei Zhang, Kevin Nguyen, Steven A Barnes, Xian-Jie Yang; Enhancing Retinal Ganglion Cell Production from Human Pluripotent Stem Cell-Derived 3D Retinal Cultures
      . Invest. Ophthalmol. Vis. Sci. 2018;59(9):564.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : In this study, we aim to determine the neurogenic potentials of the bHLH transcription factors ATOH7 and Neurog2/Ngn2, especially their abilities to influence retinal ganglion cell (RGC) development during early retinogenesis in 3D human retinal organoid cultures.

Methods : Human pluripotent stem cells were used to generate optic vesicles, which resembled undifferentiated retinal epithelium. A recombinant lentivirus encoding a Tet-inducible promoter (TetO) upstream of human ATOH7 cDNA with a C-terminal epitope tag was constructed. A similar TetO lentiviral vector encoding Neurog2/Ngn2 and the control viral vector expressing EGFP were provided by T. C. Sudhof. Concentrated lentiviral stocks were used to infect 3D optic vesicles-derived from human ES and iPS cells prior to the onset of retinogenesis and induced during the period of RGC genesis. Effects of ATOH7 and Neurog2/Ngn2 expression on RGC production were analyzed at different time intervals after Dox induction using immunocytochemistry and known RGC markers. Flow cytometry analyses were performed to quantify impacts of bHLH factor induction on neuronal differentiation. Whole cell patch clamp recording was performed to assess the function of human RGCs in adhered cultures.

Results : Lentiviral mediated expression of either bHLH neurogenic factors resulted in increased production of postmitotic neurons in 3D human retinal organoids. Quantitative analyses further revealed that induction of ATOH7 or Neurog2/Ngn2 yielded different neuronal marker profiles, suggesting distinct roles for the two factors during early retinogenesis.

Conclusions : Our results demonstrate a useful strategy to enhance human RGC production from pluripotent stem cell-derived retinal epithelium, which can serve as a model to study human retinal development and pathology.

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.

×