July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Quantitative Assessment of Retina Explant Viability in a Porcine Ex Vivo Neuroretina Model
Author Affiliations & Notes
  • Christina Lee Rettinger
    Ocular & Sensory Trauma Task Area, U.S. Army Institute of Surgical Research, Fort Sam Houston, Texas, United States
  • Heuy-Ching Hetty Wang
    Ocular & Sensory Trauma Task Area, U.S. Army Institute of Surgical Research, Fort Sam Houston, Texas, United States
  • Footnotes
    Commercial Relationships   Christina Rettinger, None; Heuy-Ching Wang, None
  • Footnotes
    Support  U.S. Army Clinical Rehabilitative Medicine Research Program, Military Operational Medicine Research Program
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 3121. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Christina Lee Rettinger, Heuy-Ching Hetty Wang; Quantitative Assessment of Retina Explant Viability in a Porcine Ex Vivo Neuroretina Model. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3121.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Ex vivo neuroretina explant cultures have been used to study an array of biological processes such as retinal development, neurodegeneration and neuroprotection. However, it is often difficult to interpret findings from these studies since most assessments used to evaluate retinal tissue behavior have largely been based on non-objective morphological measures. This prompted us to develop a technique that can assess the viability of adult retina explants quantitatively, in addition to qualitatively.

Methods : Neuroretina explants 8mm in diameter were harvested from adult swine and cultured on porous cell culture inserts with adjustable heights under serum free conditions. Retina explant viability was evaluated at 1, 4, 7, 11, 14 days of culture using a resazurin-based metabolic assay that was adapted to measure cell viability in tissue. The explants were also analyzed morphologically via immunohistochemistry for glial cell markers such as glial fibrillary acidic protein (GFAP), and terminal deoxynucleotidyl transferase-dUTP nick end labeling (TUNEL) for apoptotic cells. Moreover, neuroretina thickness was determined at each time point using hematoxylin and eosin (H&E) images. Freshly isolated, non-cultured porcine retinas were used as baseline controls.

Results : Porcine retina explants remained viable throughout the entire duration of the experiment, although the viability gradually decreased over time in culture. The laminar structure of the neuroretina was well-preserved during the first 7 days. However, by D14, most explants in culture showed significant loss of cells in each of the nuclear layers. Furthermore, there was obvious thinning of the retina explant especially in the second week of culture. Overall, the progressive loss of retinal lamination and thickness, increase in TUNEL+ nuclei with activated, hypertrophic Müller cells as evidenced by upregulated GFAP expression, were all well-correlated with the metabolic activity of the ex vivo neuroretina explants over time.

Conclusions : This study was the first report to describe the use of a high-throughput and quantitative method for monitoring retina explant viability in real-time. Even though there are limitations with these ex vivo culture systems, neuroretina cultures closely mimic the functional dynamics of the organ and provide a powerful tool that can be used to develop novel neuroprotective therapies for retinal neurodegenerative disease.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

×
×

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.

×