March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Functional Comparison Of RPE Cultures Expanded From Differentiated Human iPS Cells And Prenatal Eye Tissue
Author Affiliations & Notes
  • Ruchira Singh
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • Wei Shen
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • Xiangrong Guo
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • Enio T. Perez
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • David Kuai
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • Lynda S. Wright
    University of Wisconsin, Madison, Wisconsin
  • Bikash Pattnaik
    Depts. of Ophthalmology and Visual Sciences and Pediatrics, Eye Research Institute,
    University of Wisconsin, Madison, Wisconsin
  • David M. Gamm
    Dept. of Ophthalmology and Visual Sciences and Eye Research Institute,
    University of Wisconsin, Madison, Wisconsin
  • Footnotes
    Commercial Relationships  Ruchira Singh, None; Wei Shen, None; Xiangrong Guo, None; Enio T. Perez, None; David Kuai, None; Lynda S. Wright, None; Bikash Pattnaik, None; David M. Gamm, None
  • Footnotes
    Support  FFB Wynn-Gund Translational Research Acceleration Award; NIH R01EY21218; P30HD03352; Retina Research Foundation; UW-ICTR NIH 1UL1RR025011; UW Eye Research Institute
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 5906. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Ruchira Singh, Wei Shen, Xiangrong Guo, Enio T. Perez, David Kuai, Lynda S. Wright, Bikash Pattnaik, David M. Gamm; Functional Comparison Of RPE Cultures Expanded From Differentiated Human iPS Cells And Prenatal Eye Tissue. Invest. Ophthalmol. Vis. Sci. 2012;53(14):5906.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : Human induced pluripotent stem cells (hiPSCs) can be differentiated into retinal pigment epithelium (RPE) with varying efficiency, making hiPSCs a promising cell source for patient-specific retinal disease modeling, drug screening, and perhaps cell-based therapies. To facilitate these applications, it would be useful to isolate, passage, and expand RPE from mixed, differentiating hiPSC cultures. Therefore, we examined selected physical, molecular, and functional properties of passaged hiPSC-RPE and compared the results to those obtained from prenatal human RPE.

Methods: : hiPSC lines from two different families were differentiated toward a retinal fate using our previously described protocol. Primary hiPSC-RPE clusters were dissected, dissociated, expanded, and evaluated by RT-PCR and immunocytochemistry to assess the expression of RPE-associated genes and proteins. In addition, we examined the capacity of passaged hiPSC-RPE to form tight junctions, phagocytose bovine photoreceptor outer segments, and release calcium in response to ATP stimulation, using cultured human prenatal RPE as a control

Results: : hiPSC-RPE could be passaged at least twice and expanded >103-fold. Passaged hiPSC-RPE formed monolayers with characteristic pigmentation and morphology, expressed appropriate gene and protein markers, and formed tight junctions with transepithelial resistances (742.9 ± 37.7) on par with human prenatal RPE . Moreover, passaged hiPSC-RPE phagocytosed photoreceptor outer segments and displayed transient increases in intracellular calcium levels after ATP stimulation (89.9± 8.2 nM), similar to human prenatal RPE cultures (~100 nM).

Conclusions: : After passaging and expansion, hiPSC-RPE continued to display key cell-specific properties that were comparable to cultured human prenatal RPE. Our findings underscore the capacity of hiPSCs to serve as a versatile source of patient-specific RPE for the study and potential treatment of retinal diseases.

Keywords: retinal pigment epithelium • retinal culture • proliferation 
×
×

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.

×