June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
A microfluidic tangential flow filtration device for small EV isolation for future clinical application
Justin Leung; Dimitrios Pollalis; Chen-Ching Peng; Liya Xu; Britney Pennington; Dennis O. Clegg; Sun Young Lee
Author Affiliations & Notes
  • Justin Leung
    University of Southern California Dana and David Dornsife College of Letters Arts and Sciences, Los Angeles, California, United States
    USC Ginsburg Institute for Biomedical Therapeutics, University of Southern California, Los Angeles, California, United States
  • Dimitrios Pollalis
    USC Ginsburg Institute for Biomedical Therapeutics, University of Southern California, Los Angeles, California, United States
    Ophthalmology, USC Roski Eye Institute, University of Southern California Keck School of Medicine, Los Angeles, California, United States
  • Chen-Ching Peng
    The Vision Center at Childrens Hospital Los Angeles, Los Angeles, California, United States
  • Liya Xu
    Ophthalmology, USC Roski Eye Institute, University of Southern California Keck School of Medicine, Los Angeles, California, United States
    The Vision Center at Childrens Hospital Los Angeles, Los Angeles, California, United States
  • Britney Pennington
    Center for Stem Cell Biology and Engineering, Neuroscience Research Institute, Department of Molecular, Cellular, and Developmental Biology, University of California Santa Barbara, Santa Barbara, California, United States
  • Dennis O. Clegg
    Center for Stem Cell Biology and Engineering, Neuroscience Research Institute, Department of Molecular, Cellular, and Developmental Biology, University of California Santa Barbara, Santa Barbara, California, United States
  • Sun Young Lee
    USC Ginsburg Institute for Biomedical Therapeutics, University of Southern California, Los Angeles, California, United States
    Ophthalmology, USC Roski Eye Institute, University of Southern California Keck School of Medicine, Los Angeles, California, United States
  • Footnotes
    Commercial Relationships   Justin Leung None; Dimitrios Pollalis None; Chen-Ching Peng None; Liya Xu None; Britney Pennington None; Dennis Clegg None; Sun Young Lee None
  • Footnotes
    Support  NIH/NEI K12EY028873 and 1R56EY034193-01 to L.S.Y.; Unrestricted Grant to the Department of Ophthalmology from Research to Prevent Blindness, New York; NIH/NEI P30EY029220.
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 2741. doi:
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      Justin Leung, Dimitrios Pollalis, Chen-Ching Peng, Liya Xu, Britney Pennington, Dennis O. Clegg, Sun Young Lee; A microfluidic tangential flow filtration device for small EV isolation for future clinical application. Invest. Ophthalmol. Vis. Sci. 2023;64(8):2741.

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Abstract

Purpose : Establishing scalable and good manufacturing practice (GMP) and chemistry, manufacturing, and controls (CMC) compatible EV production is a bottleneck in developing extracellular vesicle (EV) based ocular therapeutics. To overcome this challenge, we tested a commercially available microfluidic tangential flow filtration device (mTFF) device, ExodiscTM (LabSpinner, Ulsan, South Korea), and compared it with the ultracentrifugation (UC), one of the most commonly used EV separation methods

Methods : Small EVs were separated from the cell culture supernatant (CCS) of the human embryogenic stem cell-derived fully polarized retinal pigment epithelium (hESC-PRPE-sEV) by ExodiscTM or UC. EVs were characterized by their size, numbers, morphology, tetraspanin, and multi-surface epitope heterogeneity by nanoparticle tracking analysis (NTA), TEM, Exoview, and MACSPlex Exosome Kit. Photoreceptor (PR)-preserving therapeutic efficacy of the intravitreal (IVT) injection of sEV was tested in the retinal degeneration model, RCS rats by OCT, histopathology, and ERG.

Results : The EV yield (EV particles/ml of input) and EV separation time were improved by five times in ExodiscTM compared with UC. Meanwhile, there were no significant differences in EV characteristics between the two groups, including tetraspanin heterogeneity and multiplexed phenotype. RCS rats treated with IVT injections of hESC-PRPE-sEV were able to preserve PR and their function. This therapeutic efficacy was not significantly different between sEV separated by UC and ExodiscTM.

Conclusions : Our results suggest that mTFF device holds the potential to enable large-scale and GMP and CMC-compatible EV production to develop intraocular EV therapeutics. Further studies are needed to optimize EV production, stability, and other variabilities to develop EV-derived therapeutics.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Attribution-NonCommercial-NoDerivatives
Copyright © 2024 Association for Research in Vision and Ophthalmology.
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