Bioactive hydrogels have been widely utilized for in situ tissue regeneration, promoting endogenous tissue infiltration by stimulating the host tissues. Various cell-derived extracellular matrix (cECM)-based materials have drawn attention as bioactive materials due to their bioactivity and regulating cellular function. However, the cECM-based materials still have some limitations, such as low mechanical properties and structural stability. Herein, we fabricate gelatin-based bioactive cECM (Gel-cECM) hydrogels via a thiol-ene crosslinking reaction. The cell-encapsulated Gel-cECM matrices maintain structural stability for 14 days, showing excellent cytocompatibility. Interestingly, the bioactive hydrogels promote endogenous tissue infiltration, macrophage polarization, and neovascularization when implanted in vivo. Taken together, our Gel-cECM hydrogels have great potential as a promising biomaterial for tissue regenerative medicine.