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Components:
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Lecture Sessions
• Neural stem cell biology, neurosphere and adherent monolayer culture systems
• Phenotypic and functional characterization of neural stem and progenitor cells
• Detailed technical presentation of the techniques used in neural stem cell culture systems (prior to hands-on lab work)
• Guest lecture by Dr. Brent Reynolds, Department of Neurosurgery and McKinight Brain Institute, University of Florida
• Review of current methods to identify and enumerate the neural stem cell and progenitor cells and their limitations
• Discrimination and quantification of neural stem and progenitor cells with the Neural Colony-Forming Cell (NCFC) Assay
Laboratory Sessions
• Identification of different tissue regions in the E14 and subventricular zone (SVZ) region in the adult mouse CNS
• Microdissection of tissue from embryonic and adult mouse CNS
• Dissociation techniques for CNS tissue
• Set-up of neurosphere and adherent cultures with primary mouse CNS cells
• Set-up of serum-free neuronal cultures with primary mouse cortical cells
• Calculations for cell viability, plating density and fold expansion
• Dissociation techniques for neurospheres and detachment of adherent monolayer cells
• Observation of good and poor neurosphere cultures from mouse and rat CNS cells
• Immunocytochemistry staining for the identification of neurons, astrocytes and oligodendrocytes (demonstration)
• Set-up and observation of mouse Neural Colony-Forming Cell (NCFC) Assay cultures
• Identification, quantification and isolation of colonies in the Neural Colony-Forming Cell (NCFC) Assay (demonstration)
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