Abstract
Numerous in vitro studies have been conducted in conventional static cell culture systems. However, most of the results represent an average response from a population of cells regardless of their local microenvironment. A microfluidic probe is a non-contact technology that has been widely used to perform local chemical stimulation within a restricted space, providing elaborated modulation and analysis of cellular responses within the microenvironment. Although microfluidic probes developed earlier have various potential applications, the two-dimensional structure can compromise their functionality and flexibility for practical use. In this study, we developed a three-dimensional microfluidic probe integrated device equipped with vertically oriented microchannels to overcome crucial challenges and tested the potential utility of the device in biological research. We demonstrated that the device tightly regulated spatial diffusion of a fluorescent molecule, and the flow profile predicted by simulation replicated the experimental results. Additionally, the device modulated the physiological Ca2+ response of cells within the restricted area by altering the local and temporal concentrations of biomolecules such as ATP. The novel device developed in this study may provide various applications for biological studies and contribute to further understanding of molecular mechanisms underlying cellular physiology.
Highlights
Cells communicate with one another by transmitting signals to maintain physiological homeostasis [1,2]
microfluidic probe (MFP) are simple devices used in open-culture systems like conventional cell culture dishes, the integration of MFP technology into microfluidic devices could be highly beneficial for cell-based assays
To evaluate the function of the MFP, we first performed a finite element method (FEM)-based simulation to investigate the chemical stimulation area of fluorescein, which served as a model for the fluorescence molecule
Summary
Cells communicate with one another by transmitting signals to maintain physiological homeostasis [1,2]. Several methods have been developed and employed to study the local cell-cell interactions [3,4,5,6,7,8,9,10]; it remains challenging to investigate the intercellular communication due to difficulties in spatiotemporal control of the microenvironment surrounding the cells of interest. It is important to develop methods and tools to dissect the cell-cell interactions by controlling the local microenvironment. To control the local microenvironment, a concentration gradient of injected molecules was created by gradient generators in combination with laminar flow within microchannels [11,12,13,14], and the idea was applied for high-throughput cell-based assays for biomedical research [15]. MFPs are simple devices used in open-culture systems like conventional cell culture dishes, the integration of MFP technology into microfluidic devices could be highly beneficial for cell-based assays
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