Objective: To evaluate the effect of autogenous periodontal ligament cell (PDL) transplantation in periodontal tissue regeneration. Methods: PDL cells from the same dog were cultured with a-Mem. One x 10⁵ cells of the first passage were allowed to attach to the collagen membrane for 24 hours. The membrane-cells were transplanted into a periodontal defect in the same dog. The defects were then covered with e-PTFE membranes. Defects covered with e-PTFE only; without membrane-cells, served as controls. Eighteen teeth from 6 dogs in each group were included in the study: The dogs were sacrificed at 6 weeks. Results: The results showed that new bone formation (4.00 ± 013 mm) in the test group was significantly higher than that in the control group (3.09 ± 0.28 mm, P<0.05). The new cementum formation in the test group was better than in the control group (P<0.05). Conclusion: The results suggest that PDL cell transplantation with the guided tissue regeneration technique could enhance periodontal tissue regeneration in dogs.