Core B, the Neuroscience Core, is designed to facilitate the research conducted in each of the Projects of this Program Project grant by providing assistance and support for a number of "core" activities that are common to Projects 1-4. Since neuronal stem cell-like Ntera2 cells (NT2) are used in all of the Projects, Core B will provide access to these cells as well as training in the tissue culture technology needed to induce NT2 cells to differentiate into neuron-like cells (NT2N cells) and grow up large quantities of NT2N cells. Specifically, Core B personnel will train investigators in the Projects 1-3 to induce the neuron-like NT2N cells to differentiate from the NT2 cells with retinoic acid (RA) and to generate >99% pure NT2N cells in bulk for their own studies. Since the number of NT2N cells needed fro the transplantation studies in Project 4 is modest, Core B will generate wild type and transfected NT2N cells and provide them to the investigators in Project 4. A second core activity essential to th success of Projects 1-4 is the generation, characterization and provision of antibody probes to all investigators in this Program Project. For example, well characterized antibody probes to the Beta-amyloid or A4 (BetaA4) peptide and the major amyloid precursor proteins (APPs) of the central nervous system (CNS) that are effective in Western blotting immunoprecipitation and immunohistochemical assays will be needed in each Project. Thus, Core B will be the source of such antibodies in this Program Project. Particular emphasis will be placed on identifying the epitopes in APPs recognized by new and existing monoclonal antibodies (Mabs) raised to APP isoforms expressed in E. coli or SF9 insect cells and APP synthetic peptides. Core B will provide the investigators in Projects 1-4 these anti-APP and anti-BetaA4 antibodies as well as antibodies to other proteins essential to the Projects. Finally, Core B will serve as a repository for transfected NT2/NT2N cells prepared by investigators in this Program Project and Core B will grow up these genetically modified cells on an Ad hoc basis for subsequent use in any of the Projects that require them throughout the funding period of this Program Project. In summary, by providing for several essential scientific needs of all of the Projects in this Program Project, the Neuroscience Core will play a vital role in contributing to the successful accomplishment of the research goals of this Program Project.
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