In this paper, we prepared a scaffold composed of a polyurethane (PU) fibrous outside-layer and a gelatin-heparin fibrous inner-layer with mimicking morphology and mechanical properties of a native blood vessel by sequential bilayering electrospinning technology on a rotating mandrel-type collector. The scaffolds achieved the appropriate breaking strength (3.7 ± 0.13 MPa) and elongation at break (110 ± 8%). When the scaffolds were immersed in water for 1 h, the breaking strength decreased slightly to 2.2 ± 0.3 MPa, but the elongation at break increased up to 145 ± 21%. Heparin was released from the scaffolds at substantially uniform rate until the 9th day. The scaffolds were expected to mimic the complex matrix structure of native arteries, and had good hemocompatibility as an artificial blood vessel owing to the heparin release.