Aim: The main goal of this research was to prepare a three-dimensional matrix from gingival palate tissues and investigate the possible application of this scaffold in cell culture and tissue engineering.Materials and methods: In order to fabricate the scaffolds, the biopsy samples of human palate gingival tissue were preparated surgically and divided in 5 groups then decellulization of the samples were carried out via physical method (put in nitrogen tanks and rinsing with distilled water) as well as chemical method using different concentrations of SDS (0.1%, 0.25%, 0.5%, 0.75% and 1%). Furthermore to evaluate the scaffold prepared with 1% SDS, embryonic like cells from blastema tissue were seeded on the three-dimensional scaffold.Results: Concentration of SDS below 0.5% caused significant reduction (p<0.05) of decellulization of the tissues. Microscopic studies of blastema tissue on the scaffold in different days revealed the penetration, migration, adhesion and differentiation of the cells.Conclusion: This study showed that, it is possible to prepare a natural scaffold frome palatal gingiva tissue using SDS treatment. On the other hand, the results of histologic studies showed that the decellulized scaffolds of palatal gingival might be suitable as three-dimensional bioscaffold for movement, adhesion, differentiation and migration of cells. More investigation is needed to determine the identity of the differentiated cells which further it can help to improve our knowledge about cell-matrix interaction.