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Polarized Human Retinal Pigment Epithelium Exhibits Distinct Surface Proteome on Apical and Basal Plasma Membranes

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Book cover The Surfaceome

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1722))

Abstract

Surface proteins localized on the apical and basal plasma membranes are required for a cell to sense its environment and relay changes in ionic, cytokine, chemokine, and hormone levels to the inside of the cell. In a polarized cell, surface proteins are differentially localized on the apical or the basolateral sides of the cell. The retinal pigment epithelium (RPE) is an example of a polarized cell that performs a variety of functions that are dependent on its polarized state including trafficking of ions, fluid, and metabolites across the RPE monolayer. These functions are absolutely crucial for maintaining the health and integrity of adjacent photoreceptors, the photosensitive cells of the retina. Here we present a series of approaches to identify and validate the polarization state of cultured primary human RPE cells using immunostaining for RPE apical/basolateral markers, polarized cytokine secretion, electrophysiology, fluid transport, phagocytosis, and identification of plasma membrane proteins through cell surface capturing technology. These approaches are currently being used to validate the polarized state and the epithelial phenotype of human induced pluripotent stem (iPS) cell derived RPE cells. This work provides the basis for developing an autologous cell therapy for age-related macular degeneration using patient specific iPS cell derived RPE.

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Acknowledgment

This work was supported by National Eye Institute Intramural funds.

Author information

Authors and Affiliations

  1. Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, MD, USA

    Vladimir Khristov, Qin Wan, Mostafa Lotfi & Arvydas Maminishkis

  2. Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institute of Health, Bethesda, MD, USA

    Ruchi Sharma & Kapil Bharti

Authors
  1. Vladimir Khristov
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  2. Qin Wan
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  3. Ruchi Sharma
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  4. Mostafa Lotfi
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  5. Arvydas Maminishkis
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  6. Kapil Bharti
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Corresponding author

Correspondence to Kapil Bharti .

Editor information

Editors and Affiliations

  1. School of Biomedical Sciences, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, SAR, China

    Kenneth R. Boheler

  2. Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin, USA

    Rebekah L. Gundry

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Khristov, V., Wan, Q., Sharma, R., Lotfi, M., Maminishkis, A., Bharti, K. (2018). Polarized Human Retinal Pigment Epithelium Exhibits Distinct Surface Proteome on Apical and Basal Plasma Membranes. In: Boheler, K., Gundry, R. (eds) The Surfaceome. Methods in Molecular Biology, vol 1722. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7553-2_15

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  • DOI: https://doi.org/10.1007/978-1-4939-7553-2_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7551-8

  • Online ISBN: 978-1-4939-7553-2

  • eBook Packages: Springer Protocols

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