June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
cGMP production of neural retina from hiPSCs
Author Affiliations & Notes
  • David M Gamm
    Ophthalmology and Visual Sciences, Univ of Wisconsin-Madison, Madison, WI
    Waisman Center, University of Wisconsin-Madison, Madison, WI
  • Neehar Bhatia
    Waisman Biomanufacturing, University of Wisconsin-Madison, Madison, WI
  • Anna Petelinsek
    Waisman Center, University of Wisconsin-Madison, Madison, WI
  • Jee Min
    Waisman Center, University of Wisconsin-Madison, Madison, WI
  • Elizabeth E Capowski
    Waisman Center, University of Wisconsin-Madison, Madison, WI
  • Travis Cordie
    Waisman Biomanufacturing, University of Wisconsin-Madison, Madison, WI
  • Diana Drier
    Waisman Biomanufacturing, University of Wisconsin-Madison, Madison, WI
  • Connor Lyons
    Waisman Biomanufacturing, University of Wisconsin-Madison, Madison, WI
  • Derek Hei
    Waisman Biomanufacturing, University of Wisconsin-Madison, Madison, WI
  • Joe Phillips
    Waisman Center, University of Wisconsin-Madison, Madison, WI
  • Footnotes
    Commercial Relationships David Gamm, Cellular Dynamics International (C); Neehar Bhatia, None; Anna Petelinsek, None; Jee Min, None; Elizabeth Capowski, None; Travis Cordie, None; Diana Drier, None; Connor Lyons, None; Derek Hei, None; Joe Phillips, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 3170. doi:
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    • Get Citation

      David M Gamm, Neehar Bhatia, Anna Petelinsek, Jee Min, Elizabeth E Capowski, Travis Cordie, Diana Drier, Connor Lyons, Derek Hei, Joe Phillips; cGMP production of neural retina from hiPSCs. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3170.

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

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Abstract

Purpose: Clinical trials involving human ESC and iPSC-derived RPE transplantation are well underway. However, in later stages of most outer retinal degenerative diseases, photoreceptor replacement alone or in combination with RPE must be considered. The purpose of this study is to convert an established hiPSC differentiation method for deriving neural retina (including photoreceptor precursors) to current Good Manufacturing Practices (cGMP).

Methods: All hiPSC lines investigated were created using an integration-free reprogramming method. Some hiPSC lines, including those derived from homozygous HLA “super donors,” were banked under cGMP conditions. hiPSCs were then thawed and cultured using multiple cGMP-compliant methods and differentiated to optic vesicle-like structures (OVs) using our previously established 3-D protocol. ICC analysis was performed to confirm neural retinal progenitor cell (iNRPC) and photoreceptor precursor (iPRP) cell production.

Results: Each hiPSC maintenance platform produced high, reproducible yields of OVs following retinal differentiation, with a single 6 well plate of hiPSCs giving rise to up to 400 VSX2+ OVs containing up to 24 million iNRPCs at early stages of differentiation. By day 60 of differentiation, >98% of OVs contained iPRPs as demonstrated by ICC (CRX+/RCVRN+). Cells within the OVs had >95% viability following gentle dissociation.

Conclusions: This study demonstrates robust production of hiPSC-derived neural retina across multiple cGMP-compliant culturing platforms. In our hands, E8/vitronectin is the most ideal hiPSC maintenance medium/substrate. Current studies are focused on full characterization of super donor hiPSC lines and their neural retinal derivatives and optimizing cryopreservation methods.

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