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Cartilage and osteoarthritis. Volume 1, Cellular and molecular tools / edited by Massimo Sabatini, Philippe Pastoureau, Frédéric De Ceuninck.

Contributor(s): Sabatini, Massimo | Pastoureau, Philippe | De Ceuninck, FrédéricMaterial type: TextTextSeries: Methods in molecular medicine ; 100.Publication details: Totowa, N.J. : Humana Press, 2004. Description: 1 online resource (xiv, 358 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 9781592598106; 1592598102Other title: Cellular and molecular toolsSubject(s): Cartilage cells -- Laboratory manuals | Osteoarthritis -- Laboratory manuals | Chondrocytes -- pathology | Osteoarthritis | Chondrocytes -- metabolism | HEALTH & FITNESS -- Diseases -- Musculoskeletal | Cartilage cells | OsteoarthritisGenre/Form: Electronic books. | Laboratory manuals. Additional physical formats: Print version:: Cartilage and osteoarthritis. Volume 1, Cellular and molecular tools.DDC classification: 616.7/223 LOC classification: QP88.23 | .C36eb vol. 1NLM classification: W1 | WE 348Online resources: Click here to access online
Contents:
v. 1. Culture and phenotyping of chondrocytes in primary culture -- Culture of chondrocytes in alginate beads -- Immortalization of human articular chondrocytes for generation of stable, differentiated cell lines -- Culture of immortalized chondrocytes and their use as models of chondrocyte function -- Generation of pluripotent stem cells and their differentiation to the chondrocytic phenotype -- Semiquantitative analysis of gene expression in cultured chondrocytes by RT-PCR -- Quantification of mRNA expression levels in articular chondrocytes with PCR technologies -- RNA extraction from cartilage -- Gene expression analysis in cartilage by in situ hybridization -- Analysis of differential gene expression in healthy and osteoarthritic cartilage and isolated chondrocytes by microarray analysis -- High-efficiency nonviral transfection of primary chondrocytes -- In vitro gene transfer to chondrocytes and synovial fibroblasts by adenoviral vectors -- Changes of chondrocyte metabolism in vitro: an approach by proteomic analysis -- Analysis of chondrocyte functional markers and pericellular matrix components by flow cytometry -- A simple and reliable assay of proteoglycan synthesis by cultured chondrocytes -- Assays of proteoglycan and collagen degradation in cultures of rabbit cartilage explants -- Production of antibodies against degradative neoepitopes in aggrecan -- Immunoassays for collagens in chondrocyte and cartilage explant cultures -- Detection of apoptosis in cartilage in situ and in isolated chondrocytes -- Expression, activity, and regulation of MAP kinases in cultured chondrocytes -- Mechanical loading of chondrocytes embedded in 3D constructs: in vitro methods for assessment of morphological and metabolic response to compressive strain -- In vitro physical stimulation of tissue-engineered and native cartilage.
In: Springer ProtocolsSummary: Osteoarthritis, the most common form of arthritis, imposes a significant burden of suffering on a growing population of the elderly. Even today, its poorly understood pathophysiology limits the discovery of molecular targets for pharmacological intervention and there are few effective medical treatments beyond pain control and surgery. In Cartilage and Osteoarthritis a distinguished panel of researchers, physicians, and opinion leaders in this challenging field describe their updated classical, but still evolving, techniques, as well as many emerging methods that promise to add significantly to our understanding of cartilage metabolism in health and disease. Volume 1: Cellular and Molecular Tools describes proven molecular and cellular techniques for the in vitro study of normal and osteoarthritic cartilage through biochemical, biomolecular, immunological, and physical approaches, with emphasis on the genetic manipulation of cells. The protocols follow the successful Methods in Molecular Medicine™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. A companion volume, Volume 2: Structure and In Vivo Analysis, offers cutting-edge procedures for studies-at the tissue level-of turnover, structure, and functioning in normal and diseased cartilage by invasive and noninvasive means. Comprehensive and up-to-date, the two volumes of Cartilage and Osteoarthritis provide researchers and bench scientists alike with an indispensable collection of readily reproducible protocols for new experiments-from the cellular to the animal level-designed to more clearly describe the pathophysiology of cartilage, as well as to discover novel molecular targets for pharmacological intervention.
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Includes bibliographical references and index.

v. 1. Culture and phenotyping of chondrocytes in primary culture -- Culture of chondrocytes in alginate beads -- Immortalization of human articular chondrocytes for generation of stable, differentiated cell lines -- Culture of immortalized chondrocytes and their use as models of chondrocyte function -- Generation of pluripotent stem cells and their differentiation to the chondrocytic phenotype -- Semiquantitative analysis of gene expression in cultured chondrocytes by RT-PCR -- Quantification of mRNA expression levels in articular chondrocytes with PCR technologies -- RNA extraction from cartilage -- Gene expression analysis in cartilage by in situ hybridization -- Analysis of differential gene expression in healthy and osteoarthritic cartilage and isolated chondrocytes by microarray analysis -- High-efficiency nonviral transfection of primary chondrocytes -- In vitro gene transfer to chondrocytes and synovial fibroblasts by adenoviral vectors -- Changes of chondrocyte metabolism in vitro: an approach by proteomic analysis -- Analysis of chondrocyte functional markers and pericellular matrix components by flow cytometry -- A simple and reliable assay of proteoglycan synthesis by cultured chondrocytes -- Assays of proteoglycan and collagen degradation in cultures of rabbit cartilage explants -- Production of antibodies against degradative neoepitopes in aggrecan -- Immunoassays for collagens in chondrocyte and cartilage explant cultures -- Detection of apoptosis in cartilage in situ and in isolated chondrocytes -- Expression, activity, and regulation of MAP kinases in cultured chondrocytes -- Mechanical loading of chondrocytes embedded in 3D constructs: in vitro methods for assessment of morphological and metabolic response to compressive strain -- In vitro physical stimulation of tissue-engineered and native cartilage.

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English.

Osteoarthritis, the most common form of arthritis, imposes a significant burden of suffering on a growing population of the elderly. Even today, its poorly understood pathophysiology limits the discovery of molecular targets for pharmacological intervention and there are few effective medical treatments beyond pain control and surgery. In Cartilage and Osteoarthritis a distinguished panel of researchers, physicians, and opinion leaders in this challenging field describe their updated classical, but still evolving, techniques, as well as many emerging methods that promise to add significantly to our understanding of cartilage metabolism in health and disease. Volume 1: Cellular and Molecular Tools describes proven molecular and cellular techniques for the in vitro study of normal and osteoarthritic cartilage through biochemical, biomolecular, immunological, and physical approaches, with emphasis on the genetic manipulation of cells. The protocols follow the successful Methods in Molecular Medicine™ series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. A companion volume, Volume 2: Structure and In Vivo Analysis, offers cutting-edge procedures for studies-at the tissue level-of turnover, structure, and functioning in normal and diseased cartilage by invasive and noninvasive means. Comprehensive and up-to-date, the two volumes of Cartilage and Osteoarthritis provide researchers and bench scientists alike with an indispensable collection of readily reproducible protocols for new experiments-from the cellular to the animal level-designed to more clearly describe the pathophysiology of cartilage, as well as to discover novel molecular targets for pharmacological intervention.

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