Objective To investigate the feasibility of establishing a 3D printed navigational template for cervicalpedicle screw placement, and to evaluate the accuracy of screw insertion. Methods From July 2013 to April 2015, data of 39 cases who had undergone pedicle screw placement assisted by 3D printed navigational template were retrospectively analyzed. There were 27 males and 12 females with an average age of 51.3 (range, 39-58) years old. The pathogenesis included atlantoaxial dislocation in 17 cases, cervical metastases in 8 cases, and cervical spondylosis in 14 cases. The preoperative visual analogue scale (VAS) was 7.21±3.47, and the preoperative spinal function score of Japanes Orthopaedic Association (JOA) was 10.53±2.35. All the 39 patients had undergone CT scanning before operation. After the original data was imported into Mimics 17.0 software, the 3D models of cervicalvertebrae were reconstructed.The best trajectory of pedicle screw was extracted by UGS 12.0 Imageware software according to the reverse engineering principle. According to the lamina anatomic trait, a virtual navigational template with guiding rod and hole was established. The navigational template was manufactured by 3D printing. During the operation, the navigational template was used to assist with the placement of pedicle screw. The preoperative and postoperative VAS and JOA score were compared. The accuracy of pedicle screw placement was confirmed by postoperative CT scanning. Results All the patients had been through the operations successfully. The average operation time of the patients with atlantoaxial dislocation, cervical metastases, and cervical spondylosis were 182±43 min, 217±62 min, and 235±54 min respectively, with an average blood loss of 436±287 ml, 573±291 ml, and 384±226 ml respectively. The postoperative VAS of neck pain significantly decreased to 2.91±1.24, and the post-operative JOA score was significantly increased to 14.65±2.72. 304 pedicle screws were inserted successfully, including 34 in C1, 50 in C2, 220 in C3-7. According to the Kawaguchi's standard, there were 287 screws in grade 0 (94.4%), 15 in grade 1 (4.9%), and 2 in grade 2 (0.7%). Preoperative pedicle screw transverse angle of C1, C2, and C3-7 were 8.6°±2.3°, 31.7°±8.6°, and 41.2°±9.6° respectively, and postoperative transverse angle were 9.1° ± 3.5°, 32.4° ± 7.7°, and 40.6°±9.3° respectively. Preoperative pedicle screw sagittal angle of C1, C2, and C3-7 were 9.2°±1.7°, 23.5°±4.8°, and -1.2°±2.2° respectively, and postoperative sagittal angle were 9.6°±2.1°, 22.7°±5.3°, and -1.4°±2.5° respectively. There were no significant differences between pre- and postoperative pedicle screw angles. Conclusion With the assistance of 3D printed navigational template, surgeons could change screw direction on the basis of template, clean off less soft tissues, and acquire high accuracy of cervical pedicle screw placement. Key words: Cervical vertebrae; Surgery, computer-assisted; Bone screws