# A Novel Single-Layer Microfluidic Device for Dynamic Stimulation, Culture, and Imaging of Mammalian Cells

## TL;DR

This paper introduces a new microfluidic device that allows for long-term mammalian cell culture and dynamic media mixing, enabling precise in vitro studies of cell behavior.

## Contribution

The first integration of a single-layer microfluidic mixing device with vacuum-assisted cell loading for mammalian cell culture.

## Key findings

- The device efficiently mixes and dynamically exchanges media in cell-trapping chambers.
- Mammalian cells remain viable during long-term culture in the device.
- Finite element modeling accurately predicted flow rates and mixing performance.

## Abstract

The possibility of tightly controlling the cellular microenvironment within microfluidic devices represents an important step toward precision analysis of cellular phenotypes in vitro. Microfluidic platforms that allow both long-term mammalian cell culture and dynamic modulation of the culture environment can support quantitative studies of cells’ responses to drugs. Here, we report the design and testing of a novel microfluidic device of simple production (single Polydimethylsiloxane layer), which integrates a micromixer with vacuum-assisted cell loading for long-term mammalian cell culture and dynamic mixing of four different culture media. Finite element modeling was used to predict flow rates and device dimensions to achieve diffusion-based fluid mixing. The device showed efficient mixing and dynamic exchange of media in the cell-trapping chambers, and viability of mammalian cells cultured for long-term in the device. This work represents the first attempt to integrate single-layer microfluidic mixing devices with vacuum-assisted cell-loading systems for mammalian cell culture and dynamic stimulation.