Fabrication and application of microfluidic device for 3D cell culture based on double-layer lithography

Simulating the microenvironment in vivo and constructing a neural network closer to it is of great significance to the research of the central nervous system (CNS) injury repair. Based on the above requirements, a microfluidic device for three-dimensional (3D) cell culture has been designed and verified by double-layer lithography in this paper. The device consisted of two cell chambers connected by 100 parallel microchannels which were 8 μm wide and 8 μm deep. The results of numerical simulation showed that the microfluid flow in the device was stable and uniform. 3D culture of neural stem cells (NSCs) was achieved in the device, and identified by fluorescent staining that the differentiated neuronal cell bodies were physically isolated from neurites. The device provides a dynamic, stable, and uniform microenvironment for cell culture in vitro. Together, the microfluidic device provides an important technical platform for further research on cell-cell interaction, especially axonal injury regeneration and repair.