July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Human Embryonic Stem Cells Differentiate into Trabecular Meshwork Cells
Author Affiliations & Notes
  • Yiqin Du
    Ophthalmology & Developmental Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
    McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Weitao Song
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
    Ophthalmology, Xiangya Hospital, Central South University, Changsha, Hunan, China
  • AJAY KUMAR
    Ophthalmology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Yiqin Du, None; Weitao Song, None; AJAY KUMAR, None
  • Footnotes
    Support  NIH Grant EY025643; P30-EY08098; Research to Prevent Blindness; Eye & Ear Foundation of Pittsburgh
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 5151. doi:
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      Yiqin Du, Weitao Song, AJAY KUMAR; Human Embryonic Stem Cells Differentiate into Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2019;60(9):5151.

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

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Abstract

Purpose : Stem cells for trabecular meshwork (TM) regeneration to reduce intraocular pressure is a new target for stem cell-based therapy for glaucoma. In this study, we aimed to explore a steady induction method to induce human embryonic stem cells (H9) to differentiate into TM cells fore regenerative purpose.

Methods : H9 cells were cultured and passaged on Matrigel without feeder cells. Colonies and digested monolayer cultured cells were stained with stem cell markers to confirm their stemness. The monolayer cultured H9 cells were first induced to differentiate into neural crest cells by culturing on A549 cell-derived extracellular matrix in a medium containing N2, B27 and ROCK inhibitor Y27632. Expression of neural crest cells was confirmed by flow cytometry and immunofluorescent staining. Then TM cells were induced from neural crest cells by culturing on TM cell generated extracellular matrix with a medium containing TM cell derived conditioned medium. Differentiated TM cells were confirmed by TM cell marker expression and response to dexamethasone treatment. ANOVA followed by Tukey post-test was done for statistical analysis. p < 0.05 was considered as significant.

Results : Cultured H9 colonies and monolayer cells were positive to pluripotent stem cell markers alkaline phosphatase, OCT4, SSEA4, SSEA1. The induction, the differentiated cells had reduced expression of SSEA4 and increased expression of neural crest markers NGFR and HNK1. Further induced cells started to express TM cell markers CHI3L1 and responded to dexamethasone treatment with significantly increased expression of myocilin and ANGPTL7 genes. The gene expression change was comparable to that of primary TM cells.

Conclusions : This study confirms that human embryonic stem cells can be induced to differentiate into TM cells which could be a potential source for cell-based therapy for TM regeneration for glaucoma.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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