A Y-Shaped Microfluidic Device to Study the Combined Effect of Wall Shear Stress and ATP Signals on Intracellular Calcium Dynamics in Vascular Endothelial Cells

Abstract

The intracellular calcium dynamics in vascular endothelial cells (VECs) in response to wall shear stress (WSS) and/or adenosine triphosphate (ATP) have been commonly regarded as an important factor in regulating VEC function and behavior including proliferation, migration and apoptosis. However, the effects of time-varying ATP signals have been usually neglected in the past investigations in the field of VEC mechanobiology. In order to investigate the combined effects of WSS and dynamic ATP signals on the intracellular calcium dynamic in VECs, a Y-shaped microfluidic device, which can provide the cultured cells on the bottom of its mixing micro-channel with stimuli of WSS signal alone and different combinations of WSS and ATP signals in one single micro-channel, is proposed. Both numerical simulation and experimental studies verify the feasibility of its application. Cellular experimental results also suggest that a combination of WSS and ATP signals rather than a WSS signal alone might play a more significant role in VEC Ca2+ signal transduction induced by blood flow.

A microfluidic array device for single cell capture and intracellular Ca2+ response analysis induced by dynamic biochemical stimulus

“…More specifically, as shown in Figure 3 A, the entrance of microfluidic array was connected to one syringe pump for perfusion of single cell suspension and two syringe pumps for loading dynamic biochemical stimulation which coupling by a T-bend and silicone tubes. These dynamic biochemical stimuli were generated by controlling the flow rates of the input solution and the solvent from two syringe pumps, respectively [ 25 ]. An inverted microscope (CKX41, OLYMPUS, Japan) equipped with a CCD camera (DS126431, CANON, Japan) was adopted to observe the profiles of dynamic biochemical stimuli and the intracellular Ca 2+ signal in single cells which were captured at the stagnation point of the microchannel in real time.…”

“…More specifically, as shown in Figure 3a, the entrance of microfluidic array was connected to one syringe pump for perfusion of single cell suspension and two syringe pumps for loading dynamic biochemical stimulation which coupling by a T-bend and silicone tubes. These dynamic biochemical stimuli were generated by controlling the flow rates of the input solution and the solvent from two syringe pumps, respectively [25].…”