The article reviews the state of 3D cell culture matrices, highlighting their advancements in materials chemistry, fabrication technologies, and developmental biology. These matrices better represent the geometry, chemistry, and signaling environment of natural extracellular matrix (ECM), offering significant advantages over traditional 2D cell culture methods. The review focuses on tissue engineering and in vitro modeling of human organs, emphasizing the strong potential of 3D scaffolds and cell-growth methods. Key challenges and future directions for 3D cell culture are also discussed, including the need for precise control over multi-scale structural features and the development of advanced biomaterials. The article outlines various fabrication techniques, such as electro-spinning, particulate-leaching, and solid free-form (SFF) fabrication, and their impact on matrix architecture. Additionally, it explores the selection and engineering of biomaterials, including natural and synthetic polymers, to mimic the unique characteristics of natural ECM. The review concludes by discussing the design criteria for 3D cell culture matrices, emphasizing the importance of structural and material properties, and the potential of nano-materials in biomedical engineering.The article reviews the state of 3D cell culture matrices, highlighting their advancements in materials chemistry, fabrication technologies, and developmental biology. These matrices better represent the geometry, chemistry, and signaling environment of natural extracellular matrix (ECM), offering significant advantages over traditional 2D cell culture methods. The review focuses on tissue engineering and in vitro modeling of human organs, emphasizing the strong potential of 3D scaffolds and cell-growth methods. Key challenges and future directions for 3D cell culture are also discussed, including the need for precise control over multi-scale structural features and the development of advanced biomaterials. The article outlines various fabrication techniques, such as electro-spinning, particulate-leaching, and solid free-form (SFF) fabrication, and their impact on matrix architecture. Additionally, it explores the selection and engineering of biomaterials, including natural and synthetic polymers, to mimic the unique characteristics of natural ECM. The review concludes by discussing the design criteria for 3D cell culture matrices, emphasizing the importance of structural and material properties, and the potential of nano-materials in biomedical engineering.