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Culture and Analysis of Circulating Fibrocytes

Fibrocytes circulate in the peripheral blood, produce collagen and other matrix proteins, and express cell surface markers indicative of a hematopoetic origin distinguishing them from fibroblasts. Circulating fibrocytes were first identified in 1994 in a model system of wound re ...

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Assays of T-Cell Contact Dependent Monocyte-Macrophage Functions

The production of cytokines and other inflammatory products in chronic/sterile inflammatory disorders such as rheumatoid arthritis might be induced in monocytes/macrophages by direct cellular contact with stimulated T-cells. Studies of cell-cell interactions are usual ...

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Phenotypic and Functional Analysis of Synovial Natural Killer Cells

Natural Killer (NK) cells are cells of the innate immune system with characteristic effector functions, including recognition and lysis of virus-infected or tumor cells and production of immunoregulatory cytokines, particularly interferon-gamma (IFN-γ). NK cells account for b ...

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Phenotypic Analysis of B-Cells and Plasma Cells

B-cells and antibody-secreting plasma cells are key players in protective immunity, but also in autoimmune disease. To understand their various functions in the initiation and maintenance of autoimmune pathology, a detailed dissection of their functional diversity is mandatory. ...

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Detection of Antigen Specific B-Cells in Tissues

The role of B-cells and autoantibodies in tissue destructive events of autoimmune diseases is emerging, and thereby increasing interest in identifying the presence and location of autoreactive B-cells and autoantibody secreting plasma cells. For visualization and analysis of the ...

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Single Cell Analysis of Synovial Tissue B-Cells

Mononuclear cells often form highly organized lymphoid structures in the chronically inflamed synovial tissue of patients with rheumatoid arthritis (RA) within which B-cells are activated and may differentiate into effector plasma cells. The analysis of those activated B-cells ...

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Applications of Proteomics to the Study of Adipose Tissue

Determination of the complex secretory proteome of adipocytes and its metabolic changes induced by drug treatment such as insulin or rosiglitazone is possible with the advanced proteomics technologies described herein. To study the secreted proteins of adipocytes, a 2D- liquid chr ...

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Application of DNA Microarray to the Study of Human Adipose Tissue/Cells

Adipose tissue is increasingly recognized as a metabolically active endocrine organ with multiple functions beyond its lipid storage capability. Various constituents of the tissue, such as mature adipocytes and stromal vascular cells, have distinct functions. For example, they ...

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Differentiation of Adipose Stem Cells

The broad definition of a stem cell is a cell that has the ability to self-renew and differentiate into one or more specialized terminally differentiated cell types. It has become evident that stem cells persist in, and can be isolated from, many adult tissues. Adipose tissue has been shown to contain a p ...

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Methods That Resolve Different Contributions of Clonal Expansion to Adipogenesis in 3T3-L1 and C3H10T1/2 Cells

The mouse embryo fibroblast cell lines 3T3-L1 and C3H10T1/2 differentiate to adipocytes that exhibit similar insulin regulation of lipogenesis. These cell lines, however, differ appreciably in the processes that produce the major regulator PPARγ. Each line is stimulated by a mixture of ...

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Explant Cultures of White Adipose Tissue

Obesity is characterized by increased adiposity of visceral and subcutaneous depots as well as other organs, including the vasculature. These fat depots secrete various hormone-like proteins implicated in metabolic homeostasis (e.g., adiponectin, resistin), the central cont ...

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Isolation and Culture of Preadipocytes from Rodent White Adipose Tissue

Much of the research devoted to understanding adipose tissue development is currently performed in vitro. Several cell culture models, including preadipocyte cell lines and primary culture of adipose-derived stromal vascular precursor cells, are commonly used to study molecul ...

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Flow Cytometry on the Stromal-Vascular Fraction of White Adipose Tissue

Adipose tissue contains cell types other than adipocytes that may contribute to complications linked to obesity. For example, macrophages have been shown to infiltrate adipose tissue in response to a high-fat diet. Isolation of the stromal-vascular fraction of adipose tissue allows o ...

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Application of Electrophoretic Mobility Shift Assay and Chromatin Immunoprecipitation in the Study of Transcription in Adipose Cells

Chromatin, long thought to be no more than a scaffold supporting DNA compaction inside the cell nucleus, has emerged in the last few years as a major regulatory element involved in the control of gene expression both acutely during interphase and programmatically throughout complex proces ...

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Application of RNA Interference Techniques to Adipose Cell Cultures

RNA interference (RNAi) is a powerful, quick, and easy technique to reduce the expression of a particular gene. However, investigators need to consider several steps for the experiments, including the design of the siRNA, an efficient delivery method, and a means for monitoring the biological ...

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RNA Isolation and Real-Time Quantitative RT-PCR

Adipose tissue has emerged as a major endocrine organ producing a wide spectrum of hormones and factors that play crucial roles in regulating cell turnover and function, not only locally within the adipose tissue but also in the brain and other key metabolic organ systems. It is known that gene activ ...

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Tracking Antigen Specific CD4+ T-Cells With Soluble MHC Molecules

The advent of soluble MHC multimer technology has allowed for the flow-cytometric direct identification of specific-MHC restricted antigen-specific T cells in mixed cell populations and also enabled the direct phenotyping and cloning of these cells at the same time. To date, MHC multim ...

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Identification and Manipulation of Antigen Specific T-Cells with Artificial Antigen Presenting Cells

T-cells specific for a particular antigen represent a small percentage of the overall T-cell population. Detecting the presence of antigen specific T-cells in patients, animal models or populations of cultured cells has presented a challenge to researchers. The T-cell capture method d ...

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Analysis of Antigen Reactive T-Cells

Because antigen-specific cells are the central coordinators of the immune response to infectious organisms, and the principal effector cells in autoimmune disease, there are many circumstances in which investigators may wish to examine the T-cell responses to particular antigen ...

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Analysis of Th1/Th2 T-Cell Subsets

Specific immune responses are mediated by activated CD4+ T-helper (Th) cells. Two major subsets, denoted Th1 and Th2, have been identified that are characterized by their distinctive cytokine secretion pattern and associated effector functions. The signature cytokines of Th1 and Th2 c ...

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