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细胞功能测定

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Structural Studies of MAP Kinase Cascade Components

MAPK cascade components have been the subject of structural analysis, advancing our understanding of how these enzymes are activated and how they interact. A surprising finding has been that unique inactive conformers are adopted by many of these kinases. These inactive conformers are i ...

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Adipose-Derived Stromal Cells: Cytokine Expression and Immune Cell Contaminants

The present method describes an immunoselection/depletion approach to isolate the native human adipose tissue-derived progenitor cells that are free from endothelial cells and immune cells by the use of magnetic nanobeads and microbeads coupled to antibodies. Moreover, methods ...

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Deciphering Signaling Pathways In Vivo: The Ras/Raf/Mek/Erk Cascade

The Ras/Raf/MEK/ERK cascade is a highly conserved signal transduction module, whose activation results in a number of different physiological outcomes. Depending on the cell type or the stimulus used, the pathway has been implicated in proliferation, differentiation, apoptosis, a ...

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Mutational and Functional Analysis in Human Ras/MAP Kinase Genetic Syndromes

The Ras/mitogen-activated protein kinase (MAPK) pathway is essential in regulation of the cell cycle, cell differentiation, growth, and cell senescence, each of which are critical to normal development. A class of developmental disorders, the “RASopathies,” is caused by germline mut ...

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Studies on MAP Kinase Signaling in the Immune System

The primary function of the immune system is to protect the organism from invading pathogens. In response to pathogen invasion, multiple signaling pathways are activated in immune cells, leading to diverse immune defense mechanisms. Chief among these pathways is the activation of MAPKs, ...

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Implication of the ERK Pathway on the Post-transcriptional Regulation of VEGF mRNA Stability

Vascular Endothelial Growth Factor-A (VEGF-A) is one of the most important regulators of physiological and pathological angiogenesis. Constitutive activation of the ERK pathway and over-expression of VEGF-A are common denominators of tumours of different origins. Understan ...

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Methods to Study MAP Kinase Signalling in the Central Nervous System

The mitogen-activated protein kinase (MAPK) family of intracellular signal transducers includes ERK1/2, ERK5, JNK/SAPK, and p38 and has been shown to control survival, proliferation and differentiation of cells composing the central and peripheral nervous system. Some MAPKs pre ...

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MAP Kinase Regulation of the Mitotic Spindle Checkpoint

Maintaining the integrity of the cell cycle is critical for ensuring that cells only undergo DNA replication and proliferation under controlled conditions in response to discrete stimuli. One mechanism by which the fidelity of this process is guaranteed is through the activation of cell ...

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Using High-Content Microscopy to Study Gonadotrophin-Releasing Hormone Regulation of ERK

Gonadotrophin-releasing hormone (GnRH) is a hypothalamic peptide that acts via Gq/11-coupled 7TM receptors on pituitary gonadotrophs and mediates the central control of reproduction. Recent evidence also indicates that GnRH can affect numerous tissues, but the molecular mech ...

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Isolation of Human Adipose-Derived Stem Cells from Lipoaspirates

Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Here, we describe methods from our own laboratory and the literature for the isolation and expansion of adipose-d ...

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Isolation and Culture of Rhesus Adipose-Derived Stem Cells

Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Rhesus monkeys are physiologically and phylogenetically similar to humans, and they and their cells are valua ...

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Isolation of Murine Adipose-Derived Stem Cells

Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Primary adipocytes grown in culture and derived from murine adipose tissue are essential for studying the mechanisms underlying adip ...

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In Vitro Adult Rat Adipose Tissue-Derived Stromal Cell Isolation and Differentiation

Mesenchymal stromal cell research has advanced significantly over the last decade, especially in the area of adult adipose-derived stem cells (ASCs). There are significant species differences in ASC harvest, as well as in vitro doubling time and differentiation. Selection of appropr ...

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In Vitro Adult Canine Adipose Tissue-Derived Stromal Cell Growth Characteristics

Stromal cells are undifferentiated cells found in embryonic and adult tissues. Adult mesenchymal stromal cells (MSCs) possess the properties of self renewal, long-term viability, multipotentiality, and immune privilege, which make them attractive candidates for regenerat ...

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Adipogenic Differentiation of Adult Equine Mesenchymal Stromal Cells

Equine adipose tissue-derived mesenchymal stem cells (ASCs) have only recently been investigated for their adipogenic, chondrogenic, and osteogenic differentiation potential. This chapter will briefly outline the molecular mechanisms leading to adipogenesis and the me ...

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Isolation and Culture of Porcine Adipose Tissue-Derived Somatic Stem Cells

Adipose tissue-derived stem cells (ASCs) have been described for a number of laboratory animals and humans. Improved culture conditions and cellular characteristics of ASCs have been identified. ASCs can self-renew and differentiate into multiple tissue lineages. Further chara ...

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Automated Isolation and Processing of Adipose-Derived Stem and Regenerative Cells

The popularity of nonhematopoietic, adult tissue-derived stem and progenitor cells for use as a cellular research tool, and ultimately as a clinical therapeutic, has increased exponentially over the past decade. Almost all adult-derived stem/progenitor cells (autologous and al ...

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Evaluation of Cellular and Humoral Immune Responses to Allogeneic Adipose-Derived Stem/Stromal Cells

Adipose-derived mesenchymal stem or stromal cells (ASCs) are poised for clinical use in an allogeneic setting. Although ASCs have been shown to be nonimmunogenic by several laboratories, it is advisable for the investigator to confirm this for ASCs used in their studies due to variations in ASC ...

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Characterization of Human Adipose-Derived Stem Cells Using Flow Cytometry

One of the hallmark characteristics of human adipose-derived stem cells (hASCs) is their ability to differentiate into cells of mesenchymal lineages. It is also becoming apparent that ASCs can mediate a therapeutic benefit through cytokine, paracrine-driven mechanisms influen ...

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Adipose-Derived Stem Cells for Periodontal Tissue Regeneration

Mesenchymal stem cells can effectively regenerate destroyed periodontal tissue. Because periodontal tissues are complex, mesenchymal stem cells that can differentiate into many tissue types would aid periodontal tissue regeneration. Indeed, periodontal tissue regene ...

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