The aim of this study was to determine the in vitro effect of a compound of concentrated growth factor (CGF) and periodontal ligament stem cell-derived conditioned medium (PDLSCs-CM) as a potential product for future applications in periodontal tissue regeneration. Isolated PDLSCs were characterized using flow cytometry and differentiation into osteoblasts and adipocytes cells. PDLSCs-CM and CGF were prepared and lyophilized. To determine the optimal concentration of the CGF-CM compound, the proliferation of PDLSCs after exposure to a wide range of different concentrations of CGF, CM, or their combination (CGF + CM) was investigated by methyl thiazol tetrazolium assay. Successful isolation of PDLSCs was confirmed by high expression of mesenchymal surface markers and differentiation into osteoblasts and adipocytes. PDLSCs showed higher proliferation in the 6.25 mg/mL concentration of CM and 94 μg/mL concentration of CGF. High concentrations of CGF and CM markedly inhibited the proliferation of PDLCs (p < 0.05). The exposure of PDLSCs to the compound of 10% CM + 90% CGF significantly increased the cell proliferation (p < 0.05). The results showed that CGF, CM, or their combination exert a proliferative effect on cells at a certain concentration. Further investigation on the synergistic effect of this compound may approve its application for periodontal regeneration.Keywords: Concentrated growth factor; Conditioned media; Periodontal ligament stem cells; Secretome; Tissue engineering.