This paper reports the mechanical properties of 3D layered tissue were measured by using a special fixture (micro vacuum chuck) and a measurement system. In this study, the mechanical properties of 3D layered tissue were measured by tensile test. Since difficulty of fixing the 3D layered tissue to tensile test system, the micro vacuum chuck which could fix the 3D layered tissue to tensile test system without damage was developed. Additionally, because there has been no system which can measure tensile force of 3D layered tissue, special tensile system was also developed. By using micro vacuum chuck and developed system, the tensile force of 3D layered tissue was measured and the maximum force was about about 907μN (Strain = 1.35).