Tooth autotransplantation refers to tooth transplantation from the donor to the recipient site of the same individual. Occasionally, there are situations where autotransplantation cannot be carried out immediately at clinic. In such cases, donor teeth cryopreservation is necessary to overcome limited indications. Cryopreservation media and nutrients play essential roles in preserving the periodontal ligament cells' activity. Several studies have focused on cryopreservation media and nutrients. Platelet-rich plasma (PRP) contains lots of growth factors without inducing immunologic responses or pathologic complications. Therefore, self serum and PRP might be positive factors in the cryopreservation process for preserving the periodontal ligaments (PDLs) viability. In this study, beagle dog's incisors were cryopreserved for three months in different cryopreservation media, including Dulbecco's modified eagle medium (DMEM, containing DMSO), DMEM+20% self serum, DMEM+20% self serum+PRP, and DMEM+20% fetal bovine serum (FBS). Histological results revealed that DMEM+20% self serum+PRP and DMEM+20% FBS groups showed significantly less root resorption and higher regeneration of periodontal ligament cells and that PRP is a positive factor in the cryopreservation process to preserve PDLs viability. Therefore, cryopreservation media with PRP will be a good periodontal tissue regeneration option after long-term cryopreservation in a tooth autotransplantation procedure.