<p>Regeneration of articular cartilage defects remains challenging owing to its inherently limited endogenous cell source and self-renewal capacity. Although cell therapy has shown promise in regenerating articular cartilage, maintaining cellular functional activity and precisely delivering cells to the defect site remain significant obstacles. Herein, we present a bioprinting strategy to fabricate a cartilage precursor cell (CPCs)-loaded gelatin-based scaffold, which can precisely deliver cells, maintain cellular function, and thus enhance cartilage regeneration. First, benefiting from the modification of quadruple-hydrogen-bonded ureyl pyrimidinone (UPy), gelatin modified with UPy (Gel-UPy) bioink demonstrated a temperature-programmable viscosity, enabling cell-holding stability during the printing process, and extrudable printability at a near physiological temperature. Then, through the enzymatic solidification of transglutaminase (TG), the bioprinted cell-laden scaffold (Gel-UPy@TG) was enhanced in substantial mechanical stability while maintaining a high cell survival rate. Finally, when implanted in a full-layer cartilage defect model of rabbit femoral trochlea, the bioprinted scaffold preserved the chondrogenic capacity of CPCs in the defect site and communicated with the surrounding cells, thereby accelerating the repair process of cartilage defects.</p>