June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Retinal Organoids Cultured by Microfluidic Bioreactor Demonstrated Functionality Measured by a High-Density Microelectrode Array System
Author Affiliations & Notes
  • Yuntian Xue
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
  • William C Tang
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
  • Jacqueline Chen
    Ophthalmology, University of California Irvine, Irvine, California, United States
  • Kaylee Chew
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
  • Magdalene J Seiler
    Stem Cell Research Center, University of California Irvine, Irvine, California, United States
    Physical Medicine & Rehabilitation, University of California Irvine, Irvine, California, United States
  • Andrew Browne
    Ophthalmology, University of California Irvine, Irvine, California, United States
    Biomedical Engineering, University of California Irvine, Irvine, California, United States
  • Footnotes
    Commercial Relationships   Yuntian Xue None; William Tang None; Jacqueline Chen None; Kaylee Chew None; Magdalene Seiler None; Andrew Browne None
  • Footnotes
    Support  CIRM TRAN1-10995; NIH R01 EY031834-01A1; RPB unrestricted grant to UCI Department of Ophthalmology; ICTS KL2 TR001416; Koehler Foundation Fund #6630.
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3784 – F0205. doi:
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      Yuntian Xue, William C Tang, Jacqueline Chen, Kaylee Chew, Magdalene J Seiler, Andrew Browne; Retinal Organoids Cultured by Microfluidic Bioreactor Demonstrated Functionality Measured by a High-Density Microelectrode Array System. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3784 – F0205.

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

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Abstract

Purpose : Human stem cell-derived retinal organoids (RtOgs) are able to develop matured photoreceptor cells and outer segment structures. Fully functional RtOgs should obtain complete and intact neural pathways between retinal cell layers. The purpose of this study was to apply an advanced electrophysiology testing system for verifying the functionality of matured RtOgs cultured by a customized microfluidic bioreactor platform.

Methods : RtOgs were derived from CRX-GFP genetically modified human embryonic stem cell (hESC) line (Collin et al. 2016, Stem Cells), and cultured on a customized microfluidic bioreactor system (Xue et al. 2021, Lab Chip) from day 103 to 170. On day 170, two RtOgs with outer segment structures were cut into half, with the inner layer retinal cells faced down and seeded on two high-density microelectrode arrays (HD-MEA) wells, respectively. Spontaneous discharge of RGCs and light stimulation on photoreceptor cells were measured using the full scan and network recording modes provided by the MaxOne recording system. The light stimulation experiment used natural light generated from a multi-spectral light source with controlled illuminance (ranging from 20.57 to 883.92 lux). RtOgs were cultured on the HD-MEA system from day 170 to 204 and dark-adapted before the light stimulation experiment.

Results : Spontaneous discharge of retinal cells was observed initially on day 184, the total firing rate, burst frequency, and the number of spikes per burst increased over time. RtOgs’ light responsiveness was confirmed by the denser raster scanning plot, increased firing rate, and the number of spikes per burst.

Conclusions : RtOgs cultured by the microfluidic bioreactor system developed functional inner retinal cell and photoreceptor layers, as well as the neural pathway in between. The HD-MEA recording system used in this study achieved in vitro longitudinal live recording of the RtOgs’ electrophysiological properties.

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

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