Computational Characterization of The Dish-In-A-Dish, A High Yield Culture Platform for Endothelial Shear Stress Studies on the Orbital Shaker.

Rob Driessen,Oscar Stassen,Sandra Hofmann,Carlijn Bouten,Feihu Zhao,Cecilia Sahlgren

doi:10.3390/mi11060552

Abstract

Endothelial cells sense and respond to shear stress. Different in vitro model systems have been used to study the cellular responses to shear stress, but these platforms do not allow studies on high numbers of cells under uniform and controllable shear stress. The annular dish, or dish-in-a-dish (DiaD), on the orbital shaker has been proposed as an accessible system to overcome these challenges. However, the influence of the DiaD design and the experimental parameters on the shear stress patterns is not known. In this study, we characterize different designs and experimental parameters (orbit size, speed and fluid height) using computational fluid dynamics. We optimize the DiaD for an atheroprotective flow, combining high shear stress levels with a low oscillatory shear index (OSI). We find that orbit size determines the DiaD design and parameters. The shear stress levels increase with increasing rotational speed and fluid height. Based on our optimization, we experimentally compare the 134/56 DiaD with regular dishes for cellular alignment and KLF2, eNOS, CDH2 and MCP1 expression. The calculated OSI has a strong impact on alignment and gene expression, emphasizing the importance of characterizing shear profiles in orbital setups.

Highlights

Endothelial cells line the vasculature and are sensitive to fluid shear stress induced by the blood flow
Oscillatory and low laminar (
Typical anti-inflammatory markers are Krüppel-like Factor 2 (KLF2) and endothelial nitric oxide synthase, both classical shear stress response markers found in healthy endothelium [12,13,14]

Summary

Introduction

Endothelial cells line the vasculature and are sensitive to fluid shear stress induced by the blood flow. The shear stress acting on the endothelial cells is directly linked to vascular homeostasis and disease [3,4]. Oscillatory and low laminar ( 1 Pa) are considered atheroprotective [7,10]. Cells that are exposed to an atheroprotective flow align in the direction of the flow, form a tight vascular barrier and express anti-inflammatory genes [11]. Typical anti-inflammatory markers are Krüppel-like Factor 2 (KLF2) and endothelial nitric oxide synthase (eNOS), both classical shear stress response markers found in healthy endothelium [12,13,14]

Methods

Results

Conclusion