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This detailed second edition provides key updates on well-established methods as well as coverage of recently emerging sophisticated techniques and methods that are essential for an in-depth and state-of-the-art understanding of skeletal development and homeostasis and the pathological mechanisms that cause skeletal degeneration. The first half of the book examines cell and molecular biological aspects of osteoarthritis and osteoporosis, while the remainder of the collection focuses on in vivo models of skeletal tissue injury, degeneration, and repair. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Osteoporosis and Osteoarthritis, Second Edition aims to assist investigators concerned with research topics broadly related to osteoporosis, osteoarthritis, intervertebral disc degeneration, as well as other musculoskeletal disorders.
List of contents
Isolation of Murine and Human Osteocyte.- Expansion and Chondrogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stromal Cells.- A Novel Enzymatic Digestion Approach for Isolation and Culture of Rodent Bone Marrow Mesenchymal Progenitors.- Isolation of Nucleus Pulposus and Annulus Fibrosus Cells from the Intervertebral Disc.- Engineering Cartilage Tissue by Co-Culturing of Chondrocytes and Mesenchymal Stromal Cells.- Generation of Induced Pluripotent Stem Cells.- Specimen Preparation for Single Cell Sequencing Analysis of Skeletal Cells.- Mapping 5-Hydroxymethylcytosine (5hmC) Modifications in Skeletal Tissues Using High-Throughput Sequencing.- Using FRAP to Quantify Changes in Transcription Factor Dynamics after Cell Stimulation: Cell Culture, FRAP, Data Analysis, and Visualization.- Quantitative Molecular Models for Biological Processes: Modeling of Signal Transduction Networks with ANIMO.- Generation and Characterization of Mouse Models for Skeletal Disease.- Drill Hole Models to Investigate Bone Repair.- Generation and Experimental Outcomes of Closed Femoral Fracture in Mice.- Mouse Models of Osteoarthritis: Surgical Model of Post-Traumatic Osteoarthritis Induced by Destabilization of the Medial Meniscus.- Immunostaining of Skeletal Tissues.- Mapping Regional Cortical Bone Responses to Local Changes in Loading and Systemic Stimuli.- Pain and Activity Measurements.
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