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This detailed book brings together a collection of methodologies, from the most basic to the more complex, that provides researchers with a platform they can use to embark on a cartilage research career. To aid in the search for novel therapies for cartilage regeneration, this volume addresses 3D cartilage models, challenges associated with RNA and protein extraction, imaging, gene transfer, as well as stable differentiation and variations in cell phenotype from different tissue origins. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Cartilage Tissue Engineering serves as an ideal guide for researchers working to advance the vital study of cartilage biology and repair.
List of contents
Cartilage Tissue Engineering: An Introduction.- Chondrocyte Isolation and Expansion.- Isolation of Chondrons from Hyaline Cartilage.- Articular Cartilage Chondroprogenitors: Isolation and Directed Differentiation.- Physioxic Culture of Chondrogenic Cells.- Isolation and In Vitro Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stromal Cells.- Adipose-Derived Stromal Cells: Isolation, Expansion, and Differentiation.- Chondrogenic Differentiation of Human Induced Pluripotent Stem Cells.- The Dimethylmethylene Blue Assay (DMMB) for the Quantification of Sulphated Glycosaminoglycans.- Extracellular Vesicle Isolation and Characterization for Applications in Cartilage Tissue Engineering and Osteoarthritis Therapy.- Metabolomic Profiling to Understand Chondrocyte Metabolism.- Techniques for Visualization and Quantification of Primary Cilia in Chondrocytes.- Co-Culture of Human Articular Chondrocytes Seeded in Polyurethane Scaffolds and Human Mesenchymal Stromal Cells Encapsulated in Alginate Beads.- Isolation of Murine Articular Chondrocytes for Single Cell RNA or Bulk RNA Sequencing Analysis.- Modulation of MicroRNA Expression during In Vitro Chondrogenesis.- Purification and Isolation of Proteins from Hyaline Cartilage.- Histological Preparation and Evaluation of Cartilage Specimens.- Articular Cartilage Imaging in the Context of the Superficial Chondrocyte Spatial Organization (SCSO) as a Surrogate Marker for Functional Pathology.- Unconfined Compression Experimental Protocol for Cartilage Explants and Hydrogel Constructs: From Sample Preparation to Mechanical Characterization.- Viral Gene Delivery in Chondrocytes.- Decellularization of Articular Cartilage: A Hydrochloric Acid-Based Strategy.- Self-Assembly Culture Model for Engineering Musculoskeletal Tissues.- Osteochondral Explants Isolation and Culture under a Compression and Shear Bioreactor.- A Mouse Model of Acute Cartilage Injury and Repair.- Small-Animal Compression Models of Osteoarthritis.- Modelling and Assessing Osteoarthritis in Mice by Destabilization of the Medial Meniscus (DMM).- Collagenase-Induced Osteoarthritis in Mice.- Platelet-Rich Plasma for Intra-Articular Injections: Preclinical and Clinical Evidence.
