June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Hyaluronan supports the limbal stem cell phenotype during ex vivo culture
Author Affiliations & Notes
  • Isabel Moreno
    College of Optometry, University of Houston, Houston, Texas, United States
  • Sudan Puri
    Tufts Medical Center, Boston, Massachusetts, United States
    College of Optometry, University of Houston, Houston, Texas, United States
  • Mingxia Sun
    College of Optometry, University of Houston, Houston, Texas, United States
  • Xiao Lin
    College of Optometry, University of Houston, Houston, Texas, United States
  • Tarsis F. Gesteira
    College of Optometry, University of Houston, Houston, Texas, United States
    Optimvia, Houston, Texas, United States
  • Vivien Jane Coulson-Thomas
    College of Optometry, University of Houston, Houston, Texas, United States
  • Footnotes
    Commercial Relationships   Isabel Moreno None; Sudan Puri None; Mingxia Sun None; Xiao Lin None; Tarsis Gesteira None; Vivien Coulson-Thomas None
  • Footnotes
    Support  R01 EY029289 NIH/NEI
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3657 – A0222. doi:
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    • Get Citation

      Isabel Moreno, Sudan Puri, Mingxia Sun, Xiao Lin, Tarsis F. Gesteira, Vivien Jane Coulson-Thomas; Hyaluronan supports the limbal stem cell phenotype during ex vivo culture. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3657 – A0222.

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

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Abstract

Purpose : Hyaluronan (HA), an integral component of the limbal stem cell niche (LSCN), has been shown to be instrumental in maintaining limbal stem cells (LESCs) in vivo. Here, we investigated whether human LESCs also express HA in vitro. HA is known to assist LESCs in vivo, therefore, we also investigated whether this endogenous matrix supports ex vivoexpansion of LESCs and whether the addition of exogenous HA can further support LESCs.

Methods : Primary human LESCs (hLESCs) were isolated from donor human corneas and a mouse corneal epithelial progenitor (TKE2) cell line was obtained. The HA extracellular matrix was identified in vitro through immunocytochemistry, flow cytometry, and a red blood exclusion assay. LESCs were cultured onto HA coated petri dishes or in the presence of HA through media supplementation. Cell viability, proliferation, cell size, colony formation capacity (CFC), and putative stem cell marker expression were investigated.

Results : Both hLESCs and TKE2 cells present a HA rich ECM in vitro which is essential for maintaining viable LESCs. Providing exogenous HA as supplemented media increased LESC proliferation, CFC, and putative LESC marker expression.

Conclusions : Exogenous and endogenous HA preserve the LESC phenotype supporting ex vivo expansion. The presence of HA in the ECM creates a specialized niche for LESCs both in vitro and ex vivo, therefore, supplementing hLESCs with HA can become an accessible and affordable option for clinical applications.

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

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