A pneumatic pressure-driven multi-throughput microfluidic circulation culture system

Abstract

Here, we report a pneumatic pressure-driven microfluidic device capable of multi-throughput medium circulation culture. The circulation culture system has the following advantages for application in drug discovery: (i) simultaneous operation of multiple circulation units, (ii) use of a small amount of circulating medium (3.5 mL), (iii) pipette-friendly liquid handling, and (iv) a detachable interface with pneumatic pressure lines via sterile air-vent filters. The microfluidic device contains three independent circulation culture units, in which human umbilical vein endothelial cells (HUVECs) were cultured under physiological shear stress induced by circulation of the medium. Circulation of the medium in the three culture units was generated by programmed sequentially applied pressure from two pressure-control lines. HUVECs cultured in the microfluidic device were aligned under a one-way circulating flow with a shear stress of 10 dyn cm(-2); they exhibited a randomly ordered alignment under no shear stress and under reciprocating flow with a shear stress of 10 dyn cm(-2). We also observed 2.8- to 4.9-fold increases in expression of the mRNAs of endothelial nitric oxide synthase and thrombomodulin under one-way circulating flow with a shear stress of 10 dyn cm(-2) compared with conditions of no shear stress or reciprocating flow.

Fabrication of Hollow Structures in Photodegradable Hydrogels Using a Multi-Photon Excitation Process for Blood Vessel Tissue Engineering

“…As such configuration requires complicated tube connections, it is difficult to achieve high culture throughput MPS. To solve these issues, the authors have recently developed a pressure-driven multithroughput MPS [ 16 , 51 , 52 ]. The pressure-driven MPS allows the culture solution and cells to be added to and collected from the chip by opening the lid, and further permitting medium circulation by attaching the lid and fixing it to a dedicated pneumatic attachment.…”

“…Microfluidic devices are useful tools for medium perfusion, and have recently gained immense attentions as an “organ-on-a chip” exhibiting organ level functions in vitro [ 14 ]. In the microfluidic device, vascular endothelial cells were cultured under the controlled shear stress generated by medium perfusion [ 15 , 16 ]. The flow rate can be controlled precisely even over a logarithmic range by designing the microfluidic channel network structure [ 17 , 18 ].…”

Advances in passively driven microfluidics and lab-on-chip devices: a comprehensive literature review and patent analysis

“…The authors stated that pneumatic pressure could be easily distributed to multiple wells in a reservoir with a common gas-phase space without any changes in tube connections. 134 Zhang et al developed a microuidic passive ow regulator with an in-built ve-layer structure valve for high-throughput owrate control in microuidic environments by constructing a gas-driven ow system and analyzed the ow regulation. 135 Moon et al investigated a passive microuidic ow-focusing method to generate water-in-water aqueous two-phase system (ATPS) droplets without the involvement of any external components.…”

Closed-loop feedback control for microfluidic systems through automated capacitive fluid height sensing

“…[1][2][3] Examples of mechanical pumping at these scales include syringe-pumps, microelectromechanical (MEM) pumps, 4 magnetically actuated pumps, 5,6 vacuum or pressure-driven pumps, peristaltic pumps and centrifugal pumps. 1,7,8 Chemically-induced flow can be seen in osmotic and effervescence pumps, as well as systems using electrochemical reactions. 1,[9][10][11] Surface-effect pumps include all devices making use of spontaneous capillary forces and substrate wicking to passively induce fluid flow.…”