3D cell culture is yet to be adopted and exploited to its full potential. It promises to upgrade and bring our understanding about human physiology to the highest level with the scope of applying the knowledge for better diagnosis as well as therapeutics. The focus of this book is on the direct impact of novel technologies and their evolution into viable products for the benefit of human race. It also describes the fundamentals of cell microenvironment to bring forth the relevance of 3D cell culture in tissue engineering and regenerative medicine. It discusses the extracellular matrix/microenvironment (ECM) and emphasizes its significance for growing cells in 3D to accomplish physiologically viable cell mass/tissue ex vivo. The book bridges the knowledge gaps between medical need and the technological applications through illustrations. It discusses the available models for 3D cell culture as well as the techniques to create substrates and scaffolds for achieving desired 3D microenvironment.