Objective To construct a new biomechanical testing system of in vitro spine based on LabVIEW and six-axis robot arm and conduct preliminary reliability studies, so as to make preparation for further studies on coupled motion testing. Methods By programming on LabVIEW platform, using Ethernet and Profibus to communicate the data of the robot arm with PC in real time, the displacement control and load control were realized, and the end position and rotation angle of the robot arm were read in real time. Through the force/torque sensor and data acquisition (DAQ), board DAQ, processing, storage and other functions were acquired. Using the digital micrometer and special stationary fixture, the 6-direction translation accuracy test was completed, and the data were analyzed. The flexion, extension, lateral flexion and rotation were applied on sheep spine specimen to draw the load-displacement curve, analyze and compare the experimental result, and verify the effectiveness of the biomechanical platform. Results A biomechanical test system with high precision for spine in vitro based on LabVIEW and 6-axis robot arm was developed and established, of which the average translational precision was 8.1 μm and the average translational accuracy was 56.7 μm. The program of control, data acquisition, data processing and storage were written, and the biomechanical test of the sheep spine was completed effectively. Conclusions This biomechanical test system based on Labview and 6-axis robot arm could complete the conventional three-dimensional spinal motion test with high precision and be used to conduct coupled motion test research in the next step.