In this paper, a microfluidic co-culture system comprising an embedded polydimethylsiloxane (PDMS) microstencil was fabricated. The fabricated co-culture system has two micro-channels separated with a PDMS microstencil membrane. Master molds for microchannels and stencil membranes were fabricated by photolithography, then used for casting of PDMS devices. The stencil membrane was 10 thick, with holes 10-μm large in diameter. The fabricated system co-cultured two types of cells (HepG2, NIH-3T3 Cells) successfully for seven days. The viability and stability of the cells were verified through LIVE/DEAD® staining and analysis. Additionally, albumin secretion of HepG2 cells was measured for seven days, using an HSA ELISA kit. The measured data were analyzed, to compare the activity of HepG2 cells. Results confirmed that cells can be co-cultured in the fabricated microfluidic system.