June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
IMPACT OF SUBSTRATE FLEXURAL MODULUS AND VERTICAL STABILIZATION ON PHOTRECEPTOR OUTER SEGMENT PHAGOCYTOSIS OF HUMAN IPS-DERIVED RETINAL PIGMENT EPITHELIAL CELLS
Author Affiliations & Notes
  • Tongalp H Tezel
    Ophthalmology, Columbia University, New York, New York, United States
  • Qun Zeng
    Ophthalmology, Columbia University, New York, New York, United States
  • Footnotes
    Commercial Relationships   Tongalp Tezel, None; Qun Zeng, None
  • Footnotes
    Support  Supported by an unrestricted grant from Research to Prevent Blindness, Inc, New York, NY, Foley Fund, New York, NY.
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 3285. doi:
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      Tongalp H Tezel, Qun Zeng; IMPACT OF SUBSTRATE FLEXURAL MODULUS AND VERTICAL STABILIZATION ON PHOTRECEPTOR OUTER SEGMENT PHAGOCYTOSIS OF HUMAN IPS-DERIVED RETINAL PIGMENT EPITHELIAL CELLS. Invest. Ophthalmol. Vis. Sci. 2021;62(8):3285.

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

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Abstract

Purpose : To investigate the impact of flexural modulus and vertical stabilization of the scaffolds on the ability of the human iPS-derived retinal pigment epithelial cells (hiPS-RPE) to phagocytose photoreceptor outer segments.

Methods : RPE cells from five different hiPS-RPE lines and ARPE-19 cells were cultured to confluence on human Bruch’s membrane (hBM; flexural modulus: 1.6-2.44 Mpa) explants and tissue culture inserts with a 10 µm-thick transparent polyester (PET; flexural modulus: 2.8-3.5 Gpa) membrane. One week after the confluence, 6.0 mm grafts were cut and transferred to another dish. These circular grafts were either stabilized on a photopolymerizable biogel or left unstabilized within the culture medium. Grafts were maintained within a culture medium for 4 hours in the presence of FITC-labeled human photoreceptor outer segments (108 per/ml). At the end of the incubation period, cells were collected after trypsinization. Flow cytometric analyses were carried out to determine the number of DRAQ5-stained viable cells that phagocytosed FITC-labelled human photoreceptor outer segments.

Results : hiPS-RPE exhibited better phagocytosis of photoreceptor outer segments compared to ARPE-19 cells on both substrates regardless of the graft stabilization (15.6±4.4% vs 6.8±2.4% on stabilized hBM, p=0.004; 24.1±5.5% vs 5.8±2.0% on unstabilized hBM, p=0.0001; 56.8±16.0% vs 5.7±1.6% on stabilized PET membranes, p=0.008 and 78.4±9.8% vs 29.2±8.7% on unstabilized PET membranes, p=1.5x10-6). Phagocytic ability of the hiPS-RPE cells was better if hBM (p=0.008) and PET membrane (p=4.6x10-6) were not stabilized. hiPS-RPE demonstrated better phagocytosis on PET membranes compared to hBM regardless of the stabilization of the substrate. (p<0.001)

Conclusions : Flexural modulus of the hiPS-RPE graft can modulate the function of the grafted RPE cells. hiPS-RPEcells exhibit better phagocytosis on PET membranes compared to hBM grafts.

This is a 2021 ARVO Annual Meeting abstract.

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