Development of a single layer microfluidic device for dynamic stimulation, culture and imaging of mammalian cells

The possibility to tightly control the cellular microenvironment within microfluidic devices represents an important step forward precision analysis of cellular phenotypes in vitro. In particular, microfluidic platforms that allow both long-term mammalian cell culture and dynamic modulation of the culture environment can support quantitative studies of cell responses to drugs and fate choices. Here, we report the design and testing of a novel microfluidic device of easy production (single Polydimethylsiloxane layer), which integrates a micromixer with vacuum assisted cell loading for long-term cell culture and dynamic mixing of culture media with 4 different inputs. Finite element modelling was used to predict the flow rates and device dimensions to achieve diffusion-based fluid mixing. The device showed efficient diffusion-based mixing and dynamic exchange of media in the cell trapping chambers. This work represents the first attempt to integrate single layer microfluidic mixing devices with vacuum-assisted cell loading systems for long-term mammalian cell culture and dynamic stimulation. ### Competing Interest Statement The authors have declared no competing interest.