investigations or tissue engineering require the creation of hierarchical and acellularized 3D structures mimicking the native environment of cells. Bioprinting provides a powerful approach to fabricating 3D architectures with precision and control. However, developing a bioink suitable for 3D cell culture remains challenging, particularly in achieving optimal rheological properties, printability and bioactivity necessary for cellular viability, functionality and growth. Here, we developed tissue-derived hydrogels with tunable gelation kinetics and rheological properties. By precisely adjusting the bioink's physical characteristics, we optimized its printability for extrusion-based bioprinting, enabling fast fabrication of structurally stable constructs that support the formation of 3D cellular structures. A robust decellularization protocol was developed to consistently obtain porcine skin-derived dECM (decellularized extracellular matrix) hydrogels with minimal batch-to-batch variation. The influence of dECM concentration (1-5 mg mL) on the ink's viscoelastic properties, printability, gelation kinetics, and cellular response was investigated. Gelation kinetics varied between 7 minutes to several hours, while the storage modulus ranged between 10 to 1000 Pa. Additionally, more concentrated hydrogels led to more homogeneous prints due to their higher viscosity. Fibroblast cells infiltrated the 3D matrix of the softer hydrogels (1 and 2.5 mg mL), forming an interconnected network. In contrast, migration was significantly restricted in the denser hydrogels (5 mg mL). Our findings demonstrate the potential of tissue-derived hydrogels with tunable properties for 3D bioprinting applications, enabling fast and reproducible fabrication of dECM environments for cellular studies and tissue engineering, while highlighting the critical balance between mechanical and biological properties in bioink formulation. In vitro in vivo -1 -1 -1
