This volume explores the latest collection of cell models that are used in preclinical cancer research, and covers both two-dimensional and three-dimensional culturing techniques. The chapters in this book are divided into two parts. Part One discusses two-dimensional cancer cell culture, cell models at the Air-Liquid Interface, and the latest advancements in three-dimensional complex spheroid models and dedicated disease animal models. Part Two contains technical chapters that illustrate step-by-step methodologies for specific cancer cell culture methods. The methods discussed range from the generation of isogenic cancer cell lines, the use of serum-free growth conditions, and three-dimensional cell cultures and their specific assays for the efficacy assessment of new anticancer therapies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, Cancer Cell Culture: Methods and Protocols is a valuable tool to help researchers involved in this important field to further improve or advance their models for cancer research.