Fat grafts are commonly used in plastic surgery, but their unpredictable absorption rates are a considerable disadvantage. Furthermore, no agreement has been reached regarding the method that best enables fat graft survival. This study aimed to evaluate the effects of different preparation methods on fat graft viability. Fat tissue was harvested from the remnants of transverse rectus abdominis musculocutaneous (TRAM) flaps by syringe aspiration. Harvested fat tissue was prepared using three different methods: centrifugation, metal sieve concentration, and cotton gauze concentration. To evaluate the viabilities of fat cells, XTT assays were performed. For the study, 18 nude mice were allocated to three groups: the centrifugation, metal sieve, and cotton gauze groups (6 mice per group). Prepared fat (1 ml) was injected into the nuchal area of the mice, and 12 weeks later, grafts were dissected to determine graft survival rates and subjected to histologic analysis. No significant differences were observed in graft survival rates and histologic findings (necrosis and vascularity) between the three groups. However, histologic analysis found the metal sieve group to have significantly lower fat cell viability and more inflammation than the other two groups. The findings suggest that the closed centrifugation technique has no advantage over the open cotton gauze technique in terms of fat graft viability, and that the metal sieve concentration method is deficient as a preparation method because it can cause grafted fat degradation.