June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Modeling the outer blood-retina barrier using microfluidic chips and iPSC-derived RPE and endothelial cells
Author Affiliations & Notes
  • Tea Soon Park
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Eric Nguyen
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Haig Pakhchanian
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Davide Ortolan
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Nikhil Vettikatu
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Roba Dejene
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Devika Bose
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Genqing Liang
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Ruchi Sharma
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Arvydas Maminishkis
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Kapil Bharti
    Ophthalmic Genetics & Visual Function Branch, National Eye Institute, Bethesda, Maryland, United States
  • Footnotes
    Commercial Relationships   Tea Soon Park, None; Eric Nguyen, None; Haig Pakhchanian, None; Davide Ortolan, None; Nikhil Vettikatu, None; Roba Dejene, None; Devika Bose, None; Genqing Liang, None; Ruchi Sharma, None; Arvydas Maminishkis, None; Kapil Bharti, None
  • Footnotes
    Support  NEI IRP grant ZIA EY000532-08, NEI IRP Grant ZIA EY000542-07, NEI IRP Grant ZIA EY000533-08, Bright Focus Foundation Grant M2020258
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 2192. doi:
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      Tea Soon Park, Eric Nguyen, Haig Pakhchanian, Davide Ortolan, Nikhil Vettikatu, Roba Dejene, Devika Bose, Genqing Liang, Ruchi Sharma, Arvydas Maminishkis, Kapil Bharti; Modeling the outer blood-retina barrier using microfluidic chips and iPSC-derived RPE and endothelial cells. Invest. Ophthalmol. Vis. Sci. 2021;62(8):2192.

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

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Abstract

Purpose : The retinal pigmented epithelium (RPE) and choroid vasculature form a highly specialized tissue that provides critical support for retinal function and homeostasis. Here, we combined human iPSC-derived RPE and endothelial cells (EC) with organ-on-a-chip technology, to model the outer blood-retinal barrier (oBRB). Using this platform, we aim to study patient-specific mechanisms of retinal degenerative disorders such as macular edema.

Methods : The commercially available Emulate organ-chip microfluidic system was used as the basis of the model. The apical channel of the chip was seeded with iPSC-derived RPE cells that had been differentiated for 42 days. ECs were deposited as a monolayer surrounding the chip basal channel to mimic the choroid capillaries. In order to characterize the functionality of the RPE-EC co-culture system in the chip, we assayed: (1) Dextran permeability, (2) fluid transport across apical to basal channels, and (3) RPE and EC morphology.

Results : Various genetically independent healthy and diseased iPSCs were differentiated into functional RPE and EC and cryopreserved. RPE cells formed a monolayer on the membrane of the apical channel and displayed highly pigmented polygonal morphology with expression of ZO-1 and tublin-1b. RPE tight junction integrity was confirmed using a fluorochrome conjugated dextran permeability assay, demonstrating that RPE barrier function was sufficient to block Dextran molecules. In addition, a fluid transport assay was performed to examine RPE transcytosis and homeostasis function. When compared to control chips seeded with only RPE cells, the fluid transport activity in RPE-EC chips was 2-fold higher and in range of physiological levels. These results support that both EC and RPE cells are critical components of fluid transport activity.

Conclusions : The combination of a microfluidic organ-chip system with patient-specific iPSC derivatives has provided us with a non-invasive ex vivo model to study outer retinal physiology. This micro-engineered RPE-EC chip system is a platform for the patient-specific study of diseased RPE-EC phenotypes as well as drug discovery and toxicity screening for the treatment of ocular degenerative diseases such as macular edema.

This is a 2021 ARVO Annual Meeting abstract.

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