June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Exosomes uptake by human retinal pigment epithelium cells
Author Affiliations & Notes
  • Zeljka Smit-McBride
    Ophthalmology, University of California Davis, Davis, California, United States
    Vitreoretinal Research Lab, University of California Davis, Davis, California, United States
  • Reshma Kolala
    Ophthalmology, University of California Davis, Davis, California, United States
    Vitreoretinal Research Lab, University of California Davis, Davis, California, United States
  • Ethan Lindgren
    Ophthalmology, University of California Davis, Davis, California, United States
    Vitreoretinal Research Lab, University of California Davis, Davis, California, United States
  • Ana Cristina Grodzki
    Molecular Biosciences, University of California Davis School of Veterinary Medicine, Davis, California, United States
  • Johnathon Anderson
    Otolaryngology, University of California Davis, Davis, California, United States
  • Lawrence Morse
    Ophthalmology, University of California Davis, Davis, California, United States
  • Footnotes
    Commercial Relationships   Zeljka Smit-McBride None; Reshma Kolala None; Ethan Lindgren None; Ana Cristina Grodzki None; Johnathon Anderson None; Lawrence Morse Genentech, Code C (Consultant/Contractor)
  • Footnotes
    Support  This research was supported by Barr Family Foundation, funding from an Unrestricted Departmental Grant from Research to Prevent Blindness, Inc., Dept. of Vision Science Donor Research Funds, and Mind Institute grant P50HD103526 from the National Institute of Child Health and Human Development.
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3616 – A0071. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Zeljka Smit-McBride, Reshma Kolala, Ethan Lindgren, Ana Cristina Grodzki, Johnathon Anderson, Lawrence Morse; Exosomes uptake by human retinal pigment epithelium cells. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3616 – A0071.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : The purpose of these experiments was to test and optimize uptake of mesenchymal stem cells (MSC) exosomes by human retinal pigment epithelium (ARPE-19) cell line utilizing the natural uptake mechanisms of exosomes by ARPE-19 cells.

Methods : We used human mesenchymal stem cells (MSC) exosomes labeled using ExoGlow (red-fluorescent dye) or human embryonic kidney 293 cells (HEK-293) exosomes X-Pack GFP-loaded (green-fluorescent protein). The concentrations of exosomes and the incubation period of transfected ARPE-19 were varied to determine which conditions allow for the highest transfection rate. ARPE-19 were labeled using an Oregon Green Phalloidin or CellTrace calcein red/orange AM and were imaged using an Olympus FV3000 confocal and ImageXpress Micro XL high-content microscopes. Assays were done in 96-well plates with four replicates, and plates were scanned on the ImageXpress Micro XL high-content screening system (Molecular Devices) at UC Davis IDDRC Biological Analysis Core. Additional images were obtained using a confocal microscope Olympus FV3000.

Results : Results indicate that ARPE-19 cells were able to uptake exosomes from two different heterologous sources, human mesenchymal stem cells (MSC) and human embryonic kidney 293 (HEK-293) cells. The highest transfection rate was achieved with 0.20 ug of exosomes added to 1x10^4 ARPE-19 cells/well, after a 5-hour incubation period. Imaging was best within the first 30 hours after exosomes were added to ARPE-19 cells. Signal diminished significantly after 30 hours. A higher quantity of exosomes did not equate to a higher transfection rate.

Conclusions : Exosomes are mediators of long-distance communication among different tissue and cells in the body. Exosomes carry cargo (proteins, mRNA, and microRNAs) to communicate messages to target tissues or cells. Our lab has previously identified a set of microRNAs (miRNAs) dysregulated in DR patients' ocular fluids, that may play a role in diabetic retinopathy (DR) pathogenesis. Our results strongly suggest that exosomes were quickly taken up by naïve recipient ARPE-19 cells by natural mechanisms. Our long-term goal is to develop miRNA-based therapies for DR using exosomes as natural carriers of miRNAs.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

×
×

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.

×