June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Investigating the survival and function of retinal ganglion cells in an organotypic culture: An in-vitro model for studying synaptogenesis
Author Affiliations & Notes
  • Nairouz Farah
    Life Sciences, School of Optometry, Bar Ilan University, Ramat Gan, Israel
    Bar-Ilan Institute for Nanotechnology and Advanced Materials (BINA), Bar Ilan University, Ramat Gan, Israel
  • Efrat Simon
    The Leslie and Susan Gonda Brain Research Center, Bar Ilan University, Ramat Gan, Israel
    Life Sciences, School of Optometry, Bar Ilan University, Ramat Gan, Israel
  • Yossi Mandel
    Life Sciences, School of Optometry, Bar Ilan University, Ramat Gan, Israel
    Bar-Ilan Institute for Nanotechnology and Advanced Materials (BINA), Bar Ilan University, Ramat Gan, Israel
  • Footnotes
    Commercial Relationships   Nairouz Farah None; Efrat Simon None; Yossi Mandel None
  • Footnotes
    Support  European Research Council Starter grant
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 4522 – F0309. doi:
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      Nairouz Farah, Efrat Simon, Yossi Mandel; Investigating the survival and function of retinal ganglion cells in an organotypic culture: An in-vitro model for studying synaptogenesis. Invest. Ophthalmol. Vis. Sci. 2022;63(7):4522 – F0309.

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

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Abstract

Purpose : Stem cells replacement therapy is becoming a promising pursued avenue for vision restoration in people with degenerative diseases of the outer retina. However, the integration and survival of the transplanted cells and the formation of fully functioning synapses remain a challenge. Our aim is to develop an in-vitro experimental paradigm which will allow us to address these issues while working under experimentally controlled conditions and avoiding immune system reactions faced in-vivo

Methods : As a first step, we are utilizing organotypic retinal cultures from transgenic rats expressing the calcium indicator GCaMP6f while monitoring the survival of the retinal ganglion cells (RGCs)using both extracellular recordings (multi electrode arrays), and calcium imaging at various time points.

Results : Our calcium imaging revealed robust spontaneous activity of the RGCs up to 72hrs, albeit decreasing throughout culturing period. We observe a decrease in the percentage of spontaneously active RGCs from 4.26% in acute preparations to 2.09% in 96hrs culture. Concurrently with these experiments, we have successfully established the system for extracellular investigation of RGC electrical properties incorporating flexible light pattern stimulation and multiunit analysis of the light induced responses over 60 channels. We were able to observe various RGCs types e.g., ON, OFF, ON-OFF identified by 1sec flashes applied at 0.2Hz. Moreover, through the well-known white gaussian noise stimulus combined with spike triggered averaging we generated RGCs receptive field maps obtaining the expected results both spatial (160µm FWHM) and temporally.

Conclusions : The experimental paradigm presented here can serve as a useful tool for the investigation of stem cell cells integration with the host retina, a main obstacle towards successful cell replacement based vision restoration approaches.

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

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