In this study,a stable and fast method for culturing mouse neural stem/progenitor cells (mNS/PCs) in three-dimensional (3D) type I collagen gels,based on microfluidics,is described.First,the collagen-mNS/PCs compound was driven into a microfluidic chip,and expanded with the medium consisting of DMEM/F12/RPMI1640 (1:1:1) supplemented with N2/EGF/bFGF/BSA/Lipids at a flow rate of 1μl/min.The initial medium was replaced with NB/B-27 supplemented with bFGF and BDNF when the average diameter of cell clusters was about 50-100μm at the same flow rate.The cell-collagen constructs in chip were detected by immunofluorescence test after 7 days of culture.Part of cells in the center of the 3D constructs expressed Nestin protein,and others differentiated into neurons and astrocytes,which indicates that mNS/PCs can be self-renewable and differentiate under engineered 3D microfluidics culture system.The concentration changes of Lactic acid and Glutamine in the small 3D culture system were also monitored.The concentration of lactic acid was steadily controlled within 1.0 mmol/ml,and the concentration of Glutamine was remained at around 2mmol/ml,which proves that the microfluidic cell culture system can provide a stable metabolic microenvironment for mNS/PCs 3D cultivation.In contrast,under the static culture condition,the concentration of lactic acid fluctuated between 2.5~4.5mmol/ml with batch and medium exchange modes,and the concentration of Glutamine decreased with the cells consumption and needed to be supplemented artificially.The 3D mNS/PCs model proposed here is similar to the neural tissue in morphology and cell compositions,and holds great potential for safety pharmacology,drug discovery and toxicity testing.