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丁香实验推荐阅读
A Neuron-Astrocyte Co-Culture System to Investigate Astrocyte-Secreted Factors in Mouse Neuronal Development

Astrocytes secrete factors that promote neuron survival, synapse formation, and plasticity. Understanding how these factors perform these roles requires a robust in vitro system that can effectively assess the impact of individual glial factors on neuronal properties. A classic ...

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An In Vitro Model of Reactive Astrogliosis and Its Effect on Neuronal Growth

Astrogliosis, whereby astrocytes in the central nervous system (CNS) become reactive in response to tissue damage, is a prominent process leading to the formation of the glial scar that inhibits axon regeneration after CNS injury. Upon becoming reactive, astrocytes undergo various mo ...

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Isotope-Based Quantitation of Uptake, Release, and Metabolism of Glutamate and Glucose in Cultured Astrocytes

Protocols are described for measurement in primary cultures of astrocytes of unidirectional fluxes of glutamate (influx and efflux), glutamate metabolism to glutamine or CO2, glucose influx, glycolysis, pyruvate dehydrogenation, oxidative metabolism of glucose, pyruvate ...

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Connexin-Based Channels in Astrocytes: How to Study Their Properties

A typical feature of astrocytes is their high level of connexin expression. These membrane proteins constitute the molecular basis of two types of channels: gap junction channels that allow direct cytoplasm-to-cytoplasm communication and hemichannels that provide a pathway for ex ...

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Induction of Nitric Oxide Synthase-2 Expression and Measurement of Nitric Oxide Production in Enriched Primary Cortical Astrocyte Cultures

Astrocytes produce numerous mediators under conditions of inflammation in the central nervous system. One such mediator is nitric oxide (NO) derived from nitric oxide synthase-2 (NOS-2), the high output, inducible NOS isoform. Expression of NOS-2 and production of NO can be stimulated in a ...

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ELISA Methodology to Quantify Astrocyte Production of Cytokines/Chemokines In Vitro

Astrocytes are intimately involved in immunological and inflammatory events occurring in the central nervous system (CNS), due to their ability to secrete and respond to a large number of immunoregulatory cytokines/chemokines such as IL-1β, IL-6, IL-8, IL-10, IL-17, IL-27, TNF-α, TGF-β, IFN- ...

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Use of Gel Zymography to Examine Matrix Metalloproteinase (Gelatinase) Expression in Brain Tissue or in Primary Glial Cultures

Glia synthesize, package, and secrete several species of matrix proteases, including the gelatinases (pro-)MMP-2 and (pro-)MMP-9. In appropriate settings (e.g., experimental ischemia), these MMPs can be assayed from cerebral tissues or from astrocytes and microglia in culture by enz ...

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An In Vitro Trauma Model to Study Rodent and Human Astrocyte Reactivity

Protocols are presented describing a unique in vitro injury model and how to culture and mature mouse, rat, and human astrocytes for its use. This injury model produces widespread injury and astrocyte reactivity that enable quantitative measurements of morphological, biochemical, a ...

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Astrocyte Polarization and Wound Healing in Culture: Studying Cell Adhesion Molecules

Astrocytes are highly polarized cells. This is manifest not only during development and in the adult brain, but also following injury. In response to a wound, astrocytes extend processes that participate in formation of a glial scar, which walls off lesions in the brain or spinal cord. Similarly, a ...

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Primary Cultures of Astrocytes from Fetal Bovine Brain

We describe here a method to obtain primary cell cultures from the cerebral cortex and the hypothalamus of bovine fetuses. We report how tissue origin, developmental stages, and culture medium conditions influence cell differentiation and the prevalence of glial cells vs. neurons. We com ...

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Isolation and Culture of Astrocytes from the Retina and Optic Nerve

Detailed methods for the preparation of optic nerve head (ONH) astrocytes from human donor eyes and retinal astrocytes from rat eyes are described. Included is the immunopanning method used for ONH astrocyte isolation as well as cell characterization. The isolation of purified retinal a ...

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Isolation and Culture of Spinal Cord Astrocytes

Astrocytes are possibly the most numerous cells of the vertebrate central nervous system, yet a detailed characterization of their functions is still missing. One potential reason for the obscurity of astrocytic function is that they represent a diverse population of cells that all share ...

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Derivation of Microglia-Free Astrocyte Cultures from Neural Stem Cells

Astrocytes play fundamental roles in the establishment and maintenance of tissue homeostasis in the central nervous system (CNS). To examine these different functions in vitro, it is important to be able to generate pure astrocyte cultures. While many “enriched” astrocyte cultures ha ...

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Generation of Primary Astrocyte Cultures Devoid of Contaminating Microglia

Microglia, resident phagocytic cells of the central nervous system, are frequent contaminants of astrocyte cultures. Unfortunately and not always fully appreciated, contamination by microglia can confound results of studies designed to elucidate the molecular mechanisms u ...

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Preparation of Mixed Glial Cultures from Postnatal Rat Brain

We describe a method to prepare postnatal rat brain primary cell cultures composed of astrocytes, oligodendrocytes, and microglia. After 1 week in vitro, the mixed glial cell cultures are free of neurons, meningeal cells and fibroblasts. We developed a simple procedure to selectively harv ...

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Cell Culturing of Caenorhabditis elegans Glial Cells for the Assessment of Cytosolic Ca2+ Dynamics

Cell culture has emerged as an important research method for studying various types of primary cells, including neurons and glial cells. This method has been especially instrumental in assessing intracellular Ca2+ dynamics of neural cells. The invertebrate model organism Caenorha ...

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Isolation and Culture of Human Astrocytes

Although rodent models have been essential to unveil the emerging functions of astrocytes, the existence of interspecies differences calls for caution in extrapolating data from rodent to human astrocytes. We have developed highly enriched primary astrocyte cultures from human f ...

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Isolation and Cultivation of Porcine Astrocytes

A procedure for the isolation and cultivation of astrocytes from swine is described. More specifically, the donor animals are adolescent minipigs about 3 months in age and 10 kg in weight. About 20 g of cerebral tissue can be isolated from the piglet, yielding enough astrocytes of homogeneous gene ...

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Heterogeneity of Astrocytic Form and Function

Astrocytes participate in all essential CNS functions, including blood flow regulation, energy metabolism, ion and water homeostasis, immune defence, neurotransmission, and adult neurogenesis. It is thus not surprising that astrocytic morphology and function differ betwe ...

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Neurogenic Astrocytes and Their Glycoconjugates: Not Just Glue Anymore

Cells with certain attributes of very immature astroglial cells and their radial precursors can act as stem and/or progenitor cells during developmental and persistent neurogenesis. Neural stem/progenitor cells both express and are affected by a variety of developmentally regu ...

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