〔Abstract〕 Objective To evaluate the effects and mechanisms of frozen-thawed platelet-rich plasma(PRP) stored at-80 ℃ on wound healing-related cells.Furthermore, to explore the feasibility and effectiveness of using cryopreserved PRP at low temperatures for promoting wound healing.Methods Using non-activated fresh PRP,calcium-activated fresh PRP,and post-thawed PRP,co-cultured with macrophages, fibroblasts, and vascular endothelial cells, the study measured and compared the expression of polarization and inflammatory factors associated with macrophages, as well as cell migration and proliferation rates, among other indicators, to analyze the effects of PRP on macrophage polarization, inflammation, and cell proliferation.Results Compared with the control group, post-cryopreserved PRP at ultra-low temperatures resulted in decreased expression of M1-like polarized macrophage gene iNOS( P<0. 000 1),reduced NO secretion( P<0. 001),increased urea content( P<0. 000 1),decreased M1-related inflammatory factor tumor necrosis factor-alpha( TNF-α)( P<0. 001),and decreased secretion of white blood cell interleukin( IL)-1( P<0. 001); M2-related anti-inflammatory factor IL-10 secretion levels increased( P<0. 01),and IL-12 secretion increased( P<0. 05) Furthermore,co-culturing with frozen-thawed PRP significantly promoted cell migration and enhanced vascular formation efficiency,surpassing the effects of fresh PRP and being comparable to activated PRP( P<0. 01). Cell viability assays and CCK-8 proliferation experiments also showed a significant increase in the proliferation rates of L929 and HUVEC co-cultured with frozen-thawed PRP( P<0. 01). Conclusion After being cryopreserved at-80 ℃,PRP has been proven to significantly enhance cell migration,differentiation,and proliferation capabilities,while inhibiting the production of inflammatory factors and promoting M2 polarization of macrophages. Therefore,low-temperature cryopreservation can be considered as an effective method for PRP preservation.