April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Characterization Of Phagocytic Activity In OCT4-only Derived iPS-RPE Cells
Author Affiliations & Notes
  • Stacey K. Moreno
    Cell Biology,
    The Scripps Research Institute, La Jolla, California
  • Peter D. Westenskow
    Cell Biology,
    The Scripps Research Institute, La Jolla, California
  • Tim U. Krohne
    Cell Biology,
    The Scripps Research Institute, La Jolla, California
  • Mandy Lehmann
    Cell Biology,
    The Scripps Research Institute, La Jolla, California
  • Alison L. Dorsey
    Cell Biology,
    The Scripps Research Institute, La Jolla, California
  • Saiyong Zhu
    Chemistry,
    The Scripps Research Institute, La Jolla, California
  • Sheng Ding
    Chemistry,
    The Scripps Research Institute, La Jolla, California
  • Martin Friedlander
    Cell Biology,
    The Scripps Research Institute, La Jolla, California
  • Footnotes
    Commercial Relationships  Stacey K. Moreno, None; Peter D. Westenskow, None; Tim U. Krohne, None; Mandy Lehmann, None; Alison L. Dorsey, None; Saiyong Zhu, None; Sheng Ding, None; Martin Friedlander, None
  • Footnotes
    Support  CIRM Grant TRI-01219
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 859. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Stacey K. Moreno, Peter D. Westenskow, Tim U. Krohne, Mandy Lehmann, Alison L. Dorsey, Saiyong Zhu, Sheng Ding, Martin Friedlander; Characterization Of Phagocytic Activity In OCT4-only Derived iPS-RPE Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):859.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : Retinal pigment epithelial (RPE) cells derived from induced pluripotent (iPS) cells have the potential to be used as treatment for diseases in which the native RPE are degenerated or defective such as age-related macular degeneration (AMD). In order for iPS-RPE to function as normal RPE they must have the ability to phagocytose photoreceptor outer segments (POS) in the same manner as normal RPE cells. We have developed a flow cytometry based assay to characterize phagocytosis in iPS-RPE.

Methods: : We generated iPS using retroviral delivered OCT4 and small molecules and derived RPE using directed differentiation techniques. The iPS-RPE cells strongly resemble fetal hRPE as confirmed using multiple methods. The iPS-RPE are treated with fluorescently labeled porcine outer segments over different time-points and are analyzed by flow cytometry to determine rate and efficiency of binding and internalization. Trypsin is used to release the cells from the surface and to cleave bound outer segments in order to distinguish between outer segments that are bound to the cell surface and those that are internalized. To determine receptor expression patterns during and after exposure to POS cells are labeled with antibodies to cell surface receptors thought to be involved in POS phagocytosis (such as integrin alpha v beta 5).

Results: : iPS-RPE cells phagocytose POS with slightly slower rates of binding and internalization as compared to ARPE-19 cells. Cell surface receptors involved in phagocytosis are expressed on both iPS-RPE and ARPE-19 before exposure to POS. After treating with outer segments both cell types decrease expression of integrin alpha v beta 5 on their surface and increase intracellular expression of the integrin. Other phagocytosis receptors are currently being studied.

Conclusions: : OCT4-only derived iPS-RPE express cell surface receptors known to be involved in photoreceptor phagocytosis and have the ability to phagocytose POS. Although slightly less effective as compared to cultured ARPE-19 cells iPS-RPE cells are a promising replacement for damaged RPE cells in the diseased retina.

Keywords: phagocytosis and killing • retinal pigment epithelium • flow cytometry 
×
×

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.

×