Mimicking cellular microenvironments by MEMS technology is one of the emerging research areas. Integrated biomimetic systems with nanofiber polymer networks and microfluidic chips were fabricated and cellular behaviors were observed by changing surface characteristics of nanofibers and flow rates of microchannels. Modification of polyurethane nanofiber surfaces were achieved by grafting acrylic acid with plasma treatment and these nanofiber matrices were employed in a poly(dimethylsiloxane) based microfluidic chip. The surface characteristics of both electrospun nanofiber matrices was evaluated by measuring contact angle, porosity, and chemical structure using attenuated total reflection-Fourier transform infrared spectrometry. After modification, a terminal carboxyl group formed on the nanofiber surface and the wettability increased significantly. Human MSCs were seeded on the nanofiber matrices and a morphological investigation with actin filament staining and scanning electron microscopy was performed. A proliferation test by WST-1 and Live/Dead assay were performed to investigate the cell culture environment. It was observed that the cells on the AA-grafted nanofibers spread and proliferate compared to untreated nanofibers. It has also shown that flow rates in the microchannels played an important role for cell proliferation (Sim et al., Lab Chip 2007;7:1775-1782). Integration of nanofiber matrices into the microchannels provides the useful tools for mimicking cellular microenvironments and elucidating basic questions of cell and ECM assembly and interactions.