Blood-brain barrier (BBB) is a control mechanism that limits the diffusion of many substances to central nervous system through blood, and governs the nutrient diffusivity. This barrier is among the main risk factors for treatment of neurodegenerative diseases, because most drugs designed are protein based, and are either blocked by BBB, or lose their bioavailability significantly. In this study, an in vitro BBB model was designed for testing therapeutics for neurodegenerative diseases, and methotrexate drug permeability was investigated. In the BBB model design step, polycaprolactone fiber surfaces were prepared by electrospinning to be used as support membrane for cells. The fiber morphology and sizes were determined using polarizing microscopy. Human umbilical vein endothelial cells (HUVEC) and C6 glioma cells were cultured on either side of this membrane. Model's proximity to in vivo models was tested by home-designed transendothelial electrical resistance measuring device; and nicotine was used as a positive control and albumin as a negative control. The effect of Methotrexate was determined indirectly by vitality test, MTT, for MCF-7 breast cancer cells seeded on the bottom of the well plates. The model's proximity to models in the literature and natural blood-brain barrier was specified relatively.