Objective To explore the optimal mixing ratio of advanced platelet-rich fibrin to promote the survival of fat transplantation. Methods Adipose-derived stem cells were isolated by enzymatic digestion and cultured. A-PRF was prepared from venous blood of healthy patients. A-PRF was mixed with adipose-derived stem cells at five mixing ratios of 1:6, 1:8, 1:10, 1:12, and 1:14 for wound healing assay. Sixty 6-week-old female immunodeficient mice were injected with A-PRF mixed with human fat granules at the volume ratio of 1:6, 1:8, 1:10, 1:12, and 1:14, respectively. The mice were observed continuously for twelve weeks. The volume and mass of fat samples were compared, and the density and distribution of blood vessels were observed by HE staining. The ratio of cyst/vacuole, the degree of inflammatory reaction, and the degree of interstitial fibers were analyzed. Results At 48 hours, the cell migration rate of 1:6, 1:8, 1:10, 1:12, 1:14 group were (17.67±2.49)%, (21.60±0.79)%, (24.25±0.48)%, (67.98±2.24)% and (30.55±1.18)%, respectively. At 72 hours, the migration rates of 1:6, 1:8, 1:10, 1:12, 1:14 w (30.78±1.06)%, (35.95±1.29)%, (65.47±1.64)%, (83±3.72)% and (64.12±1.99)%, respectively. The degree of cell migration in the 1:12 mixture ratio group was significantly higher than that in the other concentration groups, and the difference was statistically significant (P＜0.05 ). The retention rate of fat mass and volume in the 1:12 mixture ratio group was the highest in twelve weeks, and the difference was statistically significant (P＜0.05 ). HE staining showed that the inflammatory reaction of the five experimental groups was more severe in the first four weeks, and the inflammatory reaction was less severe in the eight and twelve weeks, and the difference was not significant. The 1:12 mixed ratio group had the lowest ratio of cyst/vacuole, the lowest degree of interstitial fibers, and a large number of new capillaries. CD31 showed that the protein content of CD31 in the 1:12 mixture ratio group was significantly higher than that in the other groups, and the difference was statistically significant (P<0.01). Conclusion The optimal mixing ratio of advanced platelet-rich fibrin to promote the survival of fat transplantation is 1:12