June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Developing iPS Cell Derived RPE Tissue for Clinical Application
Author Affiliations & Notes
  • Vladimir Khristov
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Juliet Hartford
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Qin Wan
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Mostafa Reza Lotfi
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Kiyoharu Miyagishima
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Nathan Hotaling
    Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, MD
  • Arvydas Maminishkis
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Juan Amaral
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Sheldon S Miller
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Kapil Bharti
    National Eye Institute, National Institutes of Health, Bethesda, MD
  • Footnotes
    Commercial Relationships Vladimir Khristov, None; Juliet Hartford, None; Qin Wan, None; Mostafa Lotfi, None; Kiyoharu Miyagishima, None; Nathan Hotaling, None; Arvydas Maminishkis, None; Juan Amaral, None; Sheldon Miller, None; Kapil Bharti, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1834. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Vladimir Khristov, Juliet Hartford, Qin Wan, Mostafa Reza Lotfi, Kiyoharu Miyagishima, Nathan Hotaling, Arvydas Maminishkis, Juan Amaral, Sheldon S Miller, Kapil Bharti; Developing iPS Cell Derived RPE Tissue for Clinical Application. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1834.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: Age-related macular degeneration (AMD) is a leading cause of vision loss in the United States. The disease is thought to originate by a malfunctioning retinal pigment epithelium (RPE), which leads to photoreceptor cell death and vision loss. Previous work suggests replacing diseased RPE with a healthy autologous RPE sheet can provide vision rescue for AMD patients. Here we use a tissue-engineering approach combined with induced pluripotent stem (iPS) cell technology to develop an autologous RPE sheet on an artificial biodegradable scaffold.

Methods: iPS cells are differentiated into RPE using a directed-differentiation protocol that generates RPE in three phases: neuroectoderm/RPE progenitors, committed RPE, and immature RPE. RPE cells at the immature stage are purified and seeded onto biodegradable electrospun bi-layered poly lacto-co-glycolic acid (PLGA) based scaffolds. PLGA fibers with different PLGA molecular weight are electrospun in different orientations to generate scaffolds with optimized biodegradability and tensile strength. RPE tissue is validated using electron microscopy, immunostaining, gene expression analysis, fluid transport assays, and electrophysiological analysis. RPE tissue is tested in pre-clinical animals for safety and efficacy.

Results: PLGA fiber electrospun in aligned orientation significantly increase scaffold tensile strength as compared to fibers that are electrospun in random orientation. Scaffolds with different tensile strength do not affect cell growth but affect tissue transplantation in the sub-retinal space of animal models. Electrophysiological experiments show that RPE cells on PLGA develop transepithelial resistance of more than several hundred ohms/cm2, suggesting tight electrical contacts between neighboring cells. Gene expression, immunostaining, and electron microscopy analysis show that iPS cell-derived RPE cells resemble native RPE in their molecular and structural properties. RPE cells on PLGA scaffolds also have the ability to phagocytose photoreceptor outer segments and show ability to transport water from apical to basal sides.

Conclusions: This work provides a fully-authenticated iPS cell derived RPE tissue for transplantation in pre-clinical animal models. This tissue behaves similarly to native tissue in its molecular and functional properties. This work also provides a protocol to develop a clinical-grade RPE tissue for transplantation in AMD patients.

×
×

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.

×