June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Co-Culture of Retinal Pericytes and Retinal Endothelial Cells
Author Affiliations & Notes
  • Laura Leanne Valdez
    Molecular Science, The University of Texas Rio Grande Valley School of Medicine, Edinburg, Texas, United States
  • Lili Guerra
    Molecular Science, The University of Texas Rio Grande Valley School of Medicine, Edinburg, Texas, United States
  • Richard LeBaron
    The University of Texas at San Antonio, San Antonio, Texas, United States
  • Andrew T C Tsin
    Molecular Science, The University of Texas Rio Grande Valley School of Medicine, Edinburg, Texas, United States
  • Footnotes
    Commercial Relationships   Laura Valdez None; Lili Guerra None; Richard LeBaron None; Andrew Tsin None
  • Footnotes
    Support  NIH Grants (EY033553-01; EY03355301S1 and GM137854-03)
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 1830. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Laura Leanne Valdez, Lili Guerra, Richard LeBaron, Andrew T C Tsin; Co-Culture of Retinal Pericytes and Retinal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2023;64(8):1830.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Human retinal pericytes (RP) are contractile cells adjacent to and provide support for retinal endothelial cells (REC) of retinal capillaries. One of the earliest events of diabetic retinopathy (DR) is the loss of RP through apoptosis. Pericytes share a common basement membrane with endothelial cells and are most prominent in retinal capillaries in comparison with capillaries in other tissues in the body. The loss of RP leads to the development of advanced-stage pathology including angiogenesis. In this present study of human RP and REC in coculture were examined in order to evaluate effects of their interactions and response to exogenous cytokines.

Methods : In 24-well plates a transwell insert apparatus holding 60,000 REC in the lower chamber and 40,000 RP on an upperchamber PET membrane fashioned a non-contact coculture model. Complete Classic Media (CCM) media in both chambers were changed every 24 hr. Cell number and cell viability was determined using a trypan blue dye exclusion method. Cells in each chamber well were counted twice and there was a total of two transwells per time point. In indicated experiments, REC cells were treated with 10 ug/mL of TGF-β to test for an effect on cell viability.

Results : The REC population in this co-culture system had a higher doubling time (76.6hrs) in comparison to REC grown in a single cell culture (56.4). Moreover, REC grown with RP had higher number of viable cells (175,000) compared to REC grown in a single cell culture (113,500). In contrast, RP grown in this coculture system had a decrease in doubling time (44.8 vs 76.6 hrs), along with an increase in viable cell number (229,000 vs 175,000; in comparison to cells grown in a single culture). Treatment of REC by TGF-β did not significantly alter viable cell number between coculture and single cell culture (180,000 vs 147,000).

Conclusions : We conclude that this (non-contact) coculture system of REC and RP did not significantly change growth rate nor response to TGF-β, in comparison to cell culture of a single cell type. Future study will include both contact and non-contact models to study RP and REC interactions.

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

×
×

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.

×