Background: This study aimed to compare the survival in serial autologous FAT transfer between frozen and fresh FAT among patients referred for face rejuvenation. Methods: In a clinical trial study, 60 volunteer patients for FAT injection in the nasolabial region were divided into two groups of 30. In the first group, fresh FAT was removed from the inner thigh, and 3 cc was injected on both sides of the nasolabial fold. 3 to 4 weeks later, subsequent injections were performed in the same way. In the second group, the amount of FAT required for 4 sessions of injection was removed from the inner thigh, and 3 cc was injected in the first time; the rest of the FAT was frozen at-20° C and during the next times, after defrosting in place, was injected in nasolabial region. The shelf life of adipose tissue was determined and compared between the two groups 3, 6, 9, 12, and 15 months after injection. Findings: Comparison of changes in facial wrinkle assessment score (MFWS) before and 3, 6, 9, 12, and 15 months after surgery did not show any significant difference between the two groups (P = 0. 37). The overall percentage of adipose tissue reduction during 15 months after injection was 0. 59 ± 0. 13 percent in the fresh FAT injection group and 0. 61 ± 0. 11 percent in the frozen FAT injection group with no significant difference between the two groups (P = 0. 73). Conclusion: Due to the fact that survival injected FAT is not differ between the two methods of injecting fresh and frozen FAT, the use of frozen FAT seems to be preferable for rejuvenation of the skin due to one-time FAT removal and lower operating costs.