June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Investigation of the Schlemm’s canal endothelial cells functions using a three-dimensional microfluidic model
Author Affiliations & Notes
  • Chen-Yuan Charlie Yang
    Anatomy and Neurobiology, Boston Univ School of Med, Boston, MA
    Ophthalmology, Boston University School of Medicine, Boston, MA
  • Janet Jeong
    Ophthalmology, Boston University School of Medicine, Boston, MA
  • Roger Dale Kamm
    Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA
    Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA
  • Haiyan Gong
    Anatomy and Neurobiology, Boston Univ School of Med, Boston, MA
    Ophthalmology, Boston University School of Medicine, Boston, MA
  • Footnotes
    Commercial Relationships Chen-Yuan Yang, None; Janet Jeong, None; Roger Kamm, AIM Biotech (I); Haiyan Gong, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4856. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Chen-Yuan Charlie Yang, Janet Jeong, Roger Dale Kamm, Haiyan Gong; Investigation of the Schlemm’s canal endothelial cells functions using a three-dimensional microfluidic model. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4856.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: To develop a microfluidic-based assay to study Schlemm’s canal endothelial barrier functions and giant vacuole (GV) formation under physiologically relevant basal-to-apical transendothelial flow and their responses to drug treatment.

Methods: Human Schlemm’s canal endothelial cells (HSCEC) and human dermal lymphatic microvascular endothelial cells (HLMEC) were cultured on collagen type I gel scaffold (2.5mg/mL) in microfluidic devices (based on previous design from Vickerman 2008 with modifications1). Drug-treated group was incubated with Y-27632 (25uM) for one hour prior testing for diffusive permeability (70kDa dextran) and hydraulic conductivity (9.5mmH2O basal-to-apical). Cells were perfusion-fixed and stained for F-actin. Confocal microscopy was used to visualize GVs. Permeability metrics were calculated from averaging values of individual microfluidic devices. GV numbers were determined from pooled images of all devices. Results are shown as Mean±SE.

Results: A confluent monolayer of HSCEC and HLMEC was formed and GV formation was observed when cells were exposed to a basal-to-apical flow. Y-27632 treated HSCEC showed a trend of increased diffusive permeability (2.64x10-5 ± 4.68x10-6 cm/s, n=2 vs. 1.28x10-5 ± 3.45x10-6 cm/s, n=3) and hydraulic conductivity compared to the controls. In contrast, Y-27632 treated HLMEC showed a trend of decreased diffusive permeability (1.38x10-6 ± 8.41x10-8 cm/s, n=4 vs. 2.05x10-6 ± 4.42x10-7 cm/s, n=4) and hydraulic conductivity compared to the controls. GVs exhibited the classic “signet ring” appearance; interestingly, GV numbers did not seem to differ between Y-27632 treated and control group for both HSCEC (6.5 vs. 7.5 per mm) and HLMEC (21.5 vs. 20.8 per mm).

Conclusions: We have developed a physiologically relevant in vitro three-dimensional microfluidic model to investigate Schlemm's canal endothelial barrier functions, GV formation, and their responses to drug treatment. This system creates a “trabecular outflow pathway-on-a-chip" and offers innovative ways to study cellular physiology and biomechanics of normal and glaucomatous HSCEC in the future.<br /> <br /> 1. Vickerman V, Blundo J, Chung S, Kamm R. Design, fabrication and implementation of a novel multi-parameter control microfluidic platform for three-dimensional cell culture and real-time imaging. Lab Chip 2008;8:1468-1477.

×
×

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.

×