Determination of critical shear stress for maturation of human pluripotent stem cell-derived endothelial cells towards an arterial subtype

Biotechnol Bioeng. 2019 May;116(5):1164-1175. doi: 10.1002/bit.26910. Epub 2019 Jan 23.


Human pluripotent stem cell-derived endothelial cells (hPSC-ECs) present an attractive alternative to primary EC sources for vascular grafting. However, there is a need to mature them towards either an arterial or venous subtype. A vital environmental factor involved in the arteriovenous specification of ECs during early embryonic development is fluid shear stress; therefore, there have been attempts to employ adult arterial shear stress conditions to mature hPSC-ECs. However, hPSC-ECs are naïve to fluid shear stress, and their shear responses are still not well understood. Here, we used a multiplex microfluidic platform to systematically investigate the dose-time shear responses on hPSC-EC morphology and arterial-venous phenotypes over a range of magnitudes coincidental with physiological levels of embryonic and adult vasculatures. The device comprised of six parallel cell culture chambers that were individually linked to flow-setting resistance channels, allowing us to simultaneously apply shear stress ranging from 0.4 to 15 dyne/cm 2 . We found that hPSC-ECs required up to 40 hr of shear exposure to elicit a stable phenotypic change. Cell alignment was visible at shear stress <1 dyne/cm 2 , which was independent of shear stress magnitude and duration of exposure. We discovered that the arterial markers NOTCH1 and EphrinB2 exhibited a dose-dependent increase in a similar manner beyond a threshold level of 3.8 dyne/cm 2 , whereas the venous markers COUP-TFII and EphB4 expression remained relatively constant across different magnitudes. These findings indicated that hPSC-ECs were sensitive to relatively low magnitudes of shear stress, and a critical level of ~4 dyne/cm 2 was sufficient to preferentially enhance their maturation into an arterial phenotype for future vascular tissue engineering applications.

Keywords: arterial-venous specification; biophysical cues; functional maturation; human pluripotent stem cell-derived endothelial cells; microfluidics; shear stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation*
  • Cell Line
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Ephrin-B2 / biosynthesis
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / metabolism*
  • Humans
  • Receptor, EphB4 / biosynthesis
  • Receptor, Notch1 / biosynthesis
  • Shear Strength*


  • EFNB2 protein, human
  • EPHB4 protein, human
  • Ephrin-B2
  • NOTCH1 protein, human
  • Receptor, Notch1
  • Receptor, EphB4