June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Optimized cell-handling of human embryonic stem cells in the differentiation of photoreceptor precursor cells
Author Affiliations & Notes
  • Christopher Laver
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Anat Yanai
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Aaron Joe
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Ishaq Ahmed viringipurampeer
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Xia Wang
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Andrew Metcalfe
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Cheryl Gregory-Evans
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Kevin Gregory-Evans
    Ophthalmology & Visual Sciences, University of British Columbia, Vancouver, BC, Canada
  • Footnotes
    Commercial Relationships Christopher Laver, None; Anat Yanai, None; Aaron Joe, None; Ishaq Ahmed viringipurampeer, None; Xia Wang, None; Andrew Metcalfe, None; Cheryl Gregory-Evans, None; Kevin Gregory-Evans, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1407. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Christopher Laver, Anat Yanai, Aaron Joe, Ishaq Ahmed viringipurampeer, Xia Wang, Andrew Metcalfe, Cheryl Gregory-Evans, Kevin Gregory-Evans; Optimized cell-handling of human embryonic stem cells in the differentiation of photoreceptor precursor cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1407. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: A pressing need in regenerative medicine focused on treating common blinding diseases such as age-related macular degeneration and retinitis pigmentosa is the efficient production of large numbers of human retinal precursor cells. We proposed to optimize retinal differentiation protocols for human embryonic stem cells (hESCs) by improving cell handling.

Methods: To improve efficiency we firstly focused on the production of just one retinal precursor cell type (photoreceptor precursor cells, PPCs) rather than the production of a range of retinal cells. Combining information from a number of previous studies, in particular the use of feeder-free culture media and taurine plus tri-iodothyronine supplements, we then assessed the values of using size-controlled embryoid bodies (EBs) and negative cell selection (to remove embryonic antigen-4 positive hESCs).

Results: Using size controlled 1000-cell EBs, significant improvements were made over previous studies in that 77% CRX+ve PPCs (validated via FACS) could be produced in just 17 days. This could be increased to 93% PPCs through the added step of negative cell selection. The PPC-phenotype of these cells was further supported by their expression of BLIMP1.

Conclusions: We have developed a rapid, efficient method for producing large numbers of PPCs for future preclinical studies in common blinding conditions.

Keywords: 721 stem cells • 687 regeneration • 688 retina  
×
×

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.

×