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Primary Cultures of Sympathetic Ganglia

A derivative of the neural crest, sympathetic neurons have been utilized in both in vivo and in vitro studies to approach a number of basic questions concerning the development and function of the nervous system. The superior cervical ganglia (SCG), and particularly the sympathetic chain, can ...

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Chick Spinal Somatic Motoneurons in Culture

Cultures enriched in spinal somatic motoneurons are useful test systems for delineating the role of trophic factors in motoneuron survival and differentiation (e.g., McManaman et al., 1989, McManaman et al. 1990; Juurlink et al., 1991a; Henderson et al., 1994; Mettling et al., 1995; Kaal et al., 1997). ...

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Construction and Use of Compartmented Cultures for Studies of Cell Biology of Neurons

The compartmented culture method has numerous features that make it useful for studies of nerve fiber growth, among which are: 1. Distal axons can be exposed to a different fluid and cellular environment than cell bodies and proximal axons, which is useful for studies o

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Microexplant Cultures of the Cerebellum

This chapter is devoted to a simple method for culturing developing rat cerebellum. The method can also be used for rat hippocampus embryonic age 18 d (E18), cerebral cortex (E18), and even spinal cord, but at an earlier stage of development (E14). This method was initially designed to obtain long-term ...

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Tips for Tissue Culture

Cells attach to the culture substratum with varying degrees of adherence (see Chapter 18). Cells that must attach to the substratum are referred to as “adhering cells” (anchorage-dependent cells). Cells that do not adhere and grow as cell suspensions in the medium are referred to as “nonadhering ...

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Quantification of Cells in Culture

Cell enumeration using the hemocytometer is applicable when determining the number of cells in a suspension, and when the number of samples to be analyzed is relatively small. Hemocytometry is also useful for determining the proportion of singly dispersed cells in a suspension, and for esti ...

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Elimination of Cell Types from Mixed Neural Cell Cultures

The isolation and purification, or enrichment, of neuronal or glial cell types from central nervous system (CNS) tissues require the application of one of several strategies. It is possible to separate a particular cell type, based on its cell mass or presence of specific cell surface antigens. D ...

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Colony Cultures: Plating Efficiency Assay and Cloning

The premise of colony culturing is the assumption that single viable cells can attach to the substratum, divide, and form a progeny of cells that constitute a cell colony or clone. Colony cultures can be initiated either from a disaggregated cell suspension made directly from animal tissue, or from ...

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Assay for Neuronal Cell Migration

Chemoattractant-induced neuronal migration can be directly and quantitatively evaluated in vitro, using a microchemotaxis assay. This assay has been used to quantitate the chemotropic responses of several migratory cell types, including neutrophils (Harvath et al., 1980; Gei ...

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Cultures of Astroglia and Microglia from Primary Cultures of Mouse Neopallium

Brains of newborn rats or mice are generally used as the source of tissue for glial cultures. Only about 1% of cells survive the cell disaggregation process and culture environment, and neurons that survive die within the first few days of culturing. Such cultures contain progenitor cells and glia c ...

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Preparation of Astrocyte, Oligodendrocyte, and Microglia Cultures from Primary Rat Cerebral Cultures

The study of glial cell development and function has been considerably enhanced by the development of methods to culture oligodendrocytes and astrocytes from central nervous system tissue. A primary mixed glial culture, composed mostly of astrocytes, oligodendrocytes, and micro ...

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Telomerase Inhibition and Telomere Targeting in Hematopoietic Cancer Cell Lines with Small Non-Nucleosidic Synthetic Compounds (BIBR1532)

Telomere maintenance has been shown to be essential for unlimited growth potential of human cells and is regarded as one hallmark of cancer. Telomere repeats at the ends of eukaryotic chromosomes are synthesized by the enzyme telomerase, which is active in most cancers and to some extend also in no ...

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Bovine Retinal Microvascular Pericytes: Isolation, Propagation, and Identification

The growth of new capillaries from existing vessels (angiogenesis) is of fundamental importance in wound healing and in pathological situations such as proliferative diabetic retinopathy (1), rheumatoid arthritis (2), and tumor growth. Consequently, considerable interest in ...

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Human Thymic Epithelial Cell Cultures

The thymus is a very complex organ that regulates T-cell production Thymocytes (immature T-cells) constitute by far the largest cellular population in the organ (several billions of thymocytes in a child’s thymus), but small numbers of other hematopoietic cells are found in the intrathymic ...

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Generation and Cloning of Antigen-Specific Human T-Cells

The ability to grow antigen-spectfic human T-cell clones in vitro has been instrumental in understanding T-cell function. A major breakthrough in T-cell culture in vitro was the discovery of the T-cell growth-inducing properties of interleukin-2 (IL-2), originally called T-cell growth ...

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Protection of Purified Human Hematopoietic Progenitor Cells by Interleukin-1 or Tumor Necrosis Factor-

Hematopoiesis is the process of blood cell formation in the adult human, the bone marrow (BM) is the primary hematopoietic organ. Each day, the BM produces billions of leukocytes, erythrocytes, and thrombocytes, which enter the circulation. Production of such enormous numbers of mature blo ...

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BacterialBacterial Cell Interactions in Biofilms: Detection of Polysaccharide Intercellular Adhesins by Blotting and Confocal Microscopy

Adhesive interactions between bacterial cells coupled with adherence to a solid surface can lead to the formation of a biofilm. The important role of biofilm formation in the pathogenesis of certain types of infection, especially those involving indwelling medical devices, is becomi ...

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Isolation and Culture of Murine Heart and Lung Endothelial Cells for In Vitro Model Systems

The inflammatory response is a critical component of host defense. An important goal of our group has been to understand the endothelial-dependent mechanisms that mediate leukocyte recruitment during an inflammatory response. In this chapter, we present a detailed method for the isol ...

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A Biochemical Method for Tracking Cholera Toxin Transport From Plasma Membrane to Golgi and Endoplasmic Reticulum

Asiatic cholera is a rapidly progressing disease resulting in extreme diarrhea and even death. The causative agent, cholera toxin, is an AB5 -subunit enterotoxin produced by the bacterium Vibrio cholera. The toxin must enter the intestinal cell to cause disease. Entry is achieved by the B-sub ...

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Assays for the Calcium Sensitivity of Desmosomes

Epithelial cells in vivo exist as confluent cell sheets, but this confluence is disrupted if the sheets are wounded, if the cells are undergoing morphogenesis, or if they are taking part in invasion and metastasis. Desmosomes are one of the principal types of adhesive junctions in epithelia and are ...

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