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This detailed book collects a variety of methods from investigators at the forefront of chondrocyte biology and pathology research that they have tailored for their research projects in order to tackle the unique properties and challenges of the chondrocyte and those of its tissue matrix. Divided into three parts, the volume explores experimental models to study chondrocytes, in vivo assays, as well as methods to characterize the chondrocyte phenotype with high-throughput or discrete assays on cells maintained in vivo, just isolated from their tissue (ex vivo), or cultured in vitro. Written in 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Chondrocytes: Methods andProtocols is an ideal guide for experts as well as newcomers in the cartilage field with the aim of sparking new research ideas in order to solve many of the pending mysteries regarding cartilage biology, disease mechanisms, and potential treatments.
List of contents
Isolation of Chondrocytes from Human Cartilage and Cultures in Monolayer and 3D.- Protocol for the Isolation of Intact Chondrons from Healthy and Osteoarthritic Human Articular Cartilage.- Cartilage-Specific Cre Recombinase Transgenes/Alleles in the Mouse.- Isolation of Mouse Growth Plate and Articular Chondrocytes for Primary Cultures.- Fetal Growth Plate Cartilage: Histological and Immunohistochemical Techniques.- Preparation of Adult Mouse Skeletal Tissue Sections for RNA In Situ Hybridization.- MicroRNA In Situ Hybridization in Paraffin-Embedded Human Articular Cartilage and Mouse Knee Joints.- Laser Capture Microdissection of Mouse Growth Plate Cartilage.- RNA Isolation from Articular Cartilage Tissue.- Small Nucleolar RNA Expression Profiling in Cartilage.- MicroRNA Expression Profiling, Target Identification, and Validation in Chondrocytes.- ChIP-Seq Assays from Mammalian Cartilage and Chondrocytes.- MicroRNAs and Regulation of Autophagy in Chondrocytes.- Mitochondrial Biogenesis, Activity, and DNA Isolation in Chondrocytes.- Assessing Chondrocyte Status by Immunofluorescence-Mediated Localization of Parkin Relative to Mitochondria.
