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An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment.

Publication ,  Journal Article
Cai, J; Perkumas, K; Stamer, WD; Liu, Y
Published in: Transl Vis Sci Technol
June 2020

PURPOSE: Due to the limited availability of primary human Schlemm's canal (SC) endothelial cells, we aimed to develop an in vitro cellular model using the angular aqueous plexus (AAP) cells from bovine eyes. METHODS: We harvested a mixture of cells from the trabecular meshwork region including AAP loops from multiple donors, followed by puromycin treatment and immunostaining of Von Willebrand factor and vascular endothelial (VE)-cadherin to confirm identity. Previously identified differentially expressed genes in glaucomatous SC cells were examined in non-glaucomatous SC cells (n = 3) under 0% or 15% equibiaxial strain for 24 hours using droplet digital polymerase chain reaction (ddPCR) and analyzed using the Ingenuity Pathway Analysis (IPA) software application to identify upstream regulators. To compare the cellular responses to candidate regulators of these mechanoresponsive genes, AAP and human SC cells (n = 3) were treated with 5 or 10 ng/mL transforming growth factor beta-2 (TGFβ2) for 24 or 48 hours, followed with expression profiling using real-time PCR or ddPCR. RESULTS: We found that the isolated AAP cells displayed uniform cobblestone-like morphology and positive expression of two endothelial markers. In stretched SC cells, nine glaucoma-related genes were upregulated, and IPA implicated TGFβ as a potential upstream regulator. The effects of TGFβ2 treatment were similar for both AAP and SC cells in a dose- and time-dependent manner, activating TGFBR1 and SMAD2, inhibiting BMP4, and altering expression of three glaucoma-related genes (DCN,EZR, and CYP1B1). CONCLUSIONS: Bovine AAP cells may serve as an alternative cellular model of human SC cells. TRANSLATIONAL RELEVANCE: These AAP cells may be used to study the functional pathways related to the outflow facility.

Duke Scholars

Published In

Transl Vis Sci Technol

DOI

ISSN

2164-2591

Publication Date

June 2020

Volume

9

Issue

7

Start / End Page

32

Location

United States

Related Subject Headings

  • Transforming Growth Factor beta
  • Trabecular Meshwork
  • Sclera
  • Humans
  • Endothelium
  • Endothelial Cells
  • Cattle
  • Animals
  • 3212 Ophthalmology and optometry
  • 1113 Opthalmology and Optometry
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Cai, J., Perkumas, K., Stamer, W. D., & Liu, Y. (2020). An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment. Transl Vis Sci Technol, 9(7), 32. https://doi.org/10.1167/tvst.9.7.32
Cai, Jingwen, Kristin Perkumas, W Daniel Stamer, and Yutao Liu. “An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment.Transl Vis Sci Technol 9, no. 7 (June 2020): 32. https://doi.org/10.1167/tvst.9.7.32.
Cai J, Perkumas K, Stamer WD, Liu Y. An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment. Transl Vis Sci Technol. 2020 Jun;9(7):32.
Cai, Jingwen, et al. “An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment.Transl Vis Sci Technol, vol. 9, no. 7, June 2020, p. 32. Pubmed, doi:10.1167/tvst.9.7.32.
Cai J, Perkumas K, Stamer WD, Liu Y. An In Vitro Bovine Cellular Model for Human Schlemm's Canal Endothelial Cells and Their Response to TGFβ Treatment. Transl Vis Sci Technol. 2020 Jun;9(7):32.

Published In

Transl Vis Sci Technol

DOI

ISSN

2164-2591

Publication Date

June 2020

Volume

9

Issue

7

Start / End Page

32

Location

United States

Related Subject Headings

  • Transforming Growth Factor beta
  • Trabecular Meshwork
  • Sclera
  • Humans
  • Endothelium
  • Endothelial Cells
  • Cattle
  • Animals
  • 3212 Ophthalmology and optometry
  • 1113 Opthalmology and Optometry