The　extracellular　matrix　has　drawn　considerable　interest　in　tissue　engineering　not　only　acts　as　a　bioactive　three-dimensional　scaffold　but　also　regulates　cell　behaviors　through　providing　biochemical　signals.　Extracellular　matrix-based　biomaterials,　mainly　derived　from　xenogeneic　tissues,　have　shown　positive　outcomes　in　promoting　cutaneous　wound　healing.　However,　such　extracellular　matrices　only　contain　low　doses　of　growth　factors,　which　limit　their　therapeutic　efficiency.　Recent　reports　demonstrated　that　cell　sheets　made　from　mesenchymal　stem　cell　can　accelerate　wound　repair　through　enhanced　re-epithelialization　and　angiogenesis,　but　its　clinical　translation　is　hindered　by　several　limitations,　such　as　the　risk　of　aberrant　immune　responses　and　cost　implications.　In　this　study,　acellular　extracellular　matrices　were　prepared　from　CuCl2-conditioned　mesenchymal　stem　cell　sheets　and　their　invivo　wound　healing　properties　were　evaluated　in　a　mouse　model　of　full-thickness　skin　defect.　We　found　that　extracellular　matrices　derived　from　CuCl2-conditioned　mesenchymal　stem　cell　sheets　have　a　compact　surface　with　thick　solid-like　cross-sectional　structure.　Moreover,　CuCl2　dramatically　enriched　the　extracellular　matrices　with　collagen　I,　collagen　III,　transforming　growth　factor-1,　vascular　endothelial　growth　factor,　and　basic　fibroblast　growth　factor　via　hypoxia-inducible　factor-1　activation.　And　as　a　consequence,　the　resulting　extracellular　matrices　showed　markedly　improved　invivo　wound　healing　potency　through　early　adipocyte　mobilization,　enhanced　granulation　tissues　formation,　rapid　re-epithelialization,　and　augmented　angiogenesis.　Therefore,　we　consider　that　the　extracellular　matrix　derived　from　CuCl2-conditioned　mesenchymal　stem　cell　sheets　has　the　potential　for　clinical　translation　and　may　lead　to　a　novel　strategy　for　wound　management.