3D Cell Culture Methods and Protocols

Since the publication of the previous edition, the use of 3D cell and organoid cultures has become more widespread across laboratories. This second edition volume expands on the previous edition with discussions about the latest organoid models developed for many more organs; new hydrogels and devices for 3D culture; and the organoid systems that have been improved by incorporating more components of tissue microenvironments in the in vitro culture. The chapters in this book are organized into five parts and cover topics such as biofabrication, organoids, microfluidic systems, bioprinting, and image analysis. 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.

Thorough and cutting-edge, 3D Cell Culture: Methods and Protocols, Second Edition is a valuable resource that will stimulate new ideas, foster interdisciplinary collaborations, and contribute to the improvement of human health and well-being.

3D Cell Culture: Techniques For and Beyond Organoid Applications.- Preparation of Corneal Tissue Matrix Bioink for 3D Bioprinting.- Decellularization of Pig Lung to Yield Three-Dimensional Scaffold for Lung Tissue Engineering.- Three-Dimensional Hanging Drop Spheroid Plates for Easy Chimeric Antigen Receptor (CAR) T Cytotoxicity Assay.- Static 3D Osteoblast Cell Culture on 3D Printed Titanium Scaffolds.- Modelling Acute Myeloid Leukaemia using StarPEG-Heparin Hydrogels.- A Decision Tree to Guide Human and Mouse Mammary Organoid Model Selection.- Fibroblast-Epithelium Co-Culture Methods using Epithelial Organoids and Cell Line-Derived Spheroids.- Differentiation of Fibroblasts to Adipocytes in 3D for a Co-Culture with Mammary Organoids and Immunohistological Analysis.- Organotypic 3D Cell Culture of the Embryonic Lacrimal Gland.- Renal Organoids from Whole Kidney Cells.- Organotypic 3D Ex Vivo Co-Culture Model of the Macro-Metastasis/Organ Parenchyma Interface.- Addressing the Mechanical Interaction between Cancer-Associated Fibroblasts and Cancer Cells by Laser Ablation.- Double-Barrel Perfusion System for Modification of Luminal Contents of Intestinal Organoids.- Mechanical Actuation of Organoids in Synthetic Microenvironments.- 3D Breast Cancer Model on Silk Fibroin-Integrated Microfluidic Chips.- Tumor-Microenvironment-on-Chip Platform for Assessing Drug Response in 3D Dynamic Culture.- Suspended 3D Bioprinting for Engineering Miniaturized In Vitro Tumor Models.- Real-Time Cell Cycle Imaging in a 3D Cell Culture Model of Melanoma, Quantitative Analysis, Optical Clearing, and Mathematical Modeling.- Quantitative Analysis of Whole-Mount Fluorescence-Stained Tumor Spheroids in Phenotypic Drug Screens.