Venous congestion after skin flap transplantation usually slows blood flow velocity and induces skin flap necrosis and surgical failure. Adipose tissue-derived stromal cells (ADSCs) can promote neovascularization and have been extensively applied in cell transplantation therapy and tissue regeneration. However, their function has not been reported in venous-congested skin flaps. In this study, rabbit ADSCs were isolated and identified. We established a rabbit ear venous-congested skin flap model and injected ADSCs into points along the midlines of skin flaps. The survival conditions of venous-congested skin flaps on postoperative day 7 showed that there was obvious swelling, hemorrhage, or necrosis in skin flaps of the control group, while the skin flap survival rate in the ADSC treatment group significantly increased. Hematoxylin and eosin (HE) staining results indicated that compared with the control group, thrombosis was significantly relieved and neovascularization was observed in the ADSC treatment group. Immunofluorescence revealed that the CD34 expression level and the number of capillaries significantly increased in the ADSC treatment group. In summary, ADSC transplantation promotes neovascularization in venous-congested skin flaps and skin flap survival. Therefore, ADSC transplantation may be an effective measure for promoting the survival of venous-congested skin flaps.