April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Photoreceptor- and RPE-like Cells Derived from Human Pluripotent Stem Cells Display Characteristic Responses to Small Molecule Stimulation
David M. Gamm; Jason S. Meyer; Kyle A. Wallace; Amelia D. Verhoeven; De-Ann M. Pillers; Bikash R. Pattnaik
Author Affiliations & Notes
  • David M. Gamm
    Ophthalmology and Visual Sciences, Eye Research Institute, Stem Cell Regen Med Center,
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • Jason S. Meyer
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
    Biology, IU Regen Med Center, Indiana University Purdue University Indianapolis, Indianapolis, Indiana
  • Kyle A. Wallace
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • Amelia D. Verhoeven
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • De-Ann M. Pillers
    Pediatrics,
    University of Wisconsin, Madison, Wisconsin
  • Bikash R. Pattnaik
    Pediatrics,
    Ophthalmology and Visual Sciences, Eye Research Institute,
    University of Wisconsin, Madison, Wisconsin
  • Footnotes
    Commercial Relationships  David M. Gamm, None; Jason S. Meyer, None; Kyle A. Wallace, None; Amelia D. Verhoeven, None; De-Ann M. Pillers, None; Bikash R. Pattnaik, None
  • Footnotes
    Support  FFB Wynn-Gund Translational Research Acceleration Award, Lincy Foundation, Macula Vision Research Foundation, NIH P30HD03352, Eye Research Institute, Retina Research Foundation, ICTR Pilot Grant
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 5994. doi:
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      David M. Gamm, Jason S. Meyer, Kyle A. Wallace, Amelia D. Verhoeven, De-Ann M. Pillers, Bikash R. Pattnaik; Photoreceptor- and RPE-like Cells Derived from Human Pluripotent Stem Cells Display Characteristic Responses to Small Molecule Stimulation. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5994.

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Abstract

Purpose: : To examine the physiological responses of human pluripotent stem cell (hPSC)-derived retinal cell populations following exposure to selected small molecules.

Methods: : Isolated optic vesicle(OV)-stage neurospheres derived from hPSCs were differentiated to photoreceptor(PR)- and/or RPE-like cells using an established protocol. Current responses of individual PR-like cells to applied voltage were recorded in the presence or absence of agonists and antagonists using standard whole-cell and perforated patch-clamp configurations. After recording, cells were backfilled with sulphorhodamine to facilitate identification. To monitor [Ca2+]i changes in response to ATP, hPSC-RPE cells were first loaded with Fura2AM. Thereafter, fluorescence imaging was performed to obtain time-dependent, agonist-induced [Ca2+]i measurements.

Results: : Differentiating OV neurospheres expressed appropriate gene and/or protein markers, including those involved in phototransduction. PR-like cells demonstrated an outward current at depolarizing voltages between -50 and +60 mV (holding potential = -70 mV) that was blocked by TEA. These PR-like cells possessed a resting membrane potential of -44±4 mV and a current density of 34±7.5 pA/pF at +40 mV. Upon treatment with membrane-permeable Br-cGMP, PR-like cells underwent depolarization. hPSC-RPE cells also responded to small molecule stimulation, consistently showing a transient increase in [Ca2+]i after exposure to ATP.

Conclusions: : hPSC-derived, PR- and RPE-like cells displayed important functional properties in vitro. Of particular interest, PR-like cells depolarized in the presence of the phototransduction second messenger cGMP, and RPE cells demonstrated an increase in [Ca2+]i in response to exogenous ATP, a molecule postulated to govern the RPE light response. The capacity of hPSC-derived retinal progeny to respond to physiological stimuli in vitro extends their utility as tools for basic science and clinical research.

Keywords: electrophysiology: non-clinical • photoreceptors • retinal pigment epithelium 
© 2011, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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