Bone Research Protocols
Description
This third edition volume expands on the previous editions with new chapters and updated discussions on the latest advancements in the fields of musculoskeletal research and cancer-induced bone disease (CIBD). The chapters in this book are organized in to six parts and cover a wide range of established and new research procedures. Part One looks at methods for isolation, generation and analysis of osteoclasts, stem cells, circulating tumor cells, and bone marrow adipocytes. Part Two explores biochemical and molecular analysis procedures for isolation, purification, and quantification or mRNA and DNA in bone cells. Part Three focuses on ex vivo models of tissues, organs, and co-culture systems for bone and cancer cells, and Part Four presents various cancer related in vivo models of primary bone and secondary cancers in the skeleton. Part 5 discusses the frequently used bone microscopical and imaging analytical techniques, such as bone histomorphometry, immunostaining, and MicroCT scanning of bone. Finally Part Six talks about applications of GWAS, EWAS, systematic review, and meta-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 trouble3.shooting and avoiding known pitfalls.
Cutting-edge and comprehensive,
Bone Research Protocols, Third Edition
is a valuable resource for all researchers, scientists, and clinicians who are interested in learning more about this important and developing field.
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Content
Isolation and Culture of Osteoblasts.- Isolation and Generation of Osteoclasts.- Isolation, Purification, Generation, and Culture of Osteocytes.- Isolation, Differentiation, and Characterization of Human Bone Marrow Stromal Stem Cells In Vitro and In Vivo.- Isolation and Characterization of Primary Bone Marrow Adipocytes in Rodent Models.- Isolation and Characterization of Circulating Tumor Cells.- 2D and 3D In Vitro Co-Culture for Cancer and Bone Cell Interaction Studies.- Isolation of Extracellular Vesicle Subtypes from Primary Osteoblast Cultures.- Using Cell and Organ Culture Models to Analyze Responses of Bone Cells to Mechanical Stimulation.- Analysis of Signaling Pathways by Western Blotting and Immunoprecipitation.- Analysis of mRNA, miRNA, and DNA in Bone Cells by RT-qPCR and In Situ Hybridization.- Analysis of Transcriptional Regulation in Bone Cells.- The Ex Vivo Organ Culture of Bone and Cartilage.- Ex Vivo Model Systems of Cancer-Bone Cell Interactions.- Mechanical Properties of Bone Ex Vivo.- Ovariectomy/Orchidectomy in Rodents.- Osteoarthritis Mouse Model of Destabilization of the Medial Meniscus.- Models of Rheumatoid Arthritis.- In Vivo Models of Mechanical Loading.- Pre-Clinical Murine Models of Bone Sarcomas.- Mouse Models of Breast Cancer Bone Metastasis.- Radiographic Assessment of Osteosclerotic Lesions in Mice with Bone Metastasis.- Histomorphometry in Rodents.- Immunostaining of Skeletal Tissues.- Live Cell Imaging of Bone Cell and Organ Cultures.- Analysis of Bone Architecture in Rodents Using Micro Computed Tomography.- 3D Morphometry of Preclinical Bone Disease Models Beyond Osteoporosis.- MicroCT Scanning and Analysis in the Zebrafish Skeleton.- Ex Vivo Examining Single Cancer Cells Homing to Murine Bone Using Two-Photon Microscopy.- Quantitative X-Ray Imaging of Mouse Bone by Faxitron.- Scanning Electron Microscopy and Bone.- Fourier Transform Infrared Imaging of Bone.- Raman Microscopy and Bone.- Analysis of Genome-Wide Association Studies.- Analysis of Epigenome-Wide Association Studies.- Application of Systematic Review and Meta-Analysis in Musculoskeletal Research.