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Cryopreservation of Cortical Tissue Blocks for the Generation of Highly Enriched Neuronal Cultures
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November 11, 2010
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Here, we describe a method for efficient cryopreservation and thawing of cortical brain tissue blocks to generate highly enriched neuronal cultures. This simple protocol provides flexibility for later generation of neuronal, astrocyte, and neuronal precursor cell cultures.
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CryopreservationNeuronal CulturesFreezing MediumDimethyl Sulfoxide (DMSO)Hank’s Buffered Salt Solution (HBSS)CryovialsRate-controlled Freezing ContainerPost-thaw ProcessingDissociationNeuronal-enriched CulturesNeuritic GrowthImmunocytochemical StainingAstrocytic MarkerGlial Fibrillary Acidic Protein (GFAP)Neuronal MarkerBeta-tubulin Class IIINeural Precursor Cell CulturesNeurospheresDifferentiating Conditions
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Rahman, A. S., Parvinjah, S., Hanna, M. A., Helguera, P. R., Busciglio, J. Cryopreservation of Cortical Tissue Blocks for the Generation of Highly Enriched Neuronal Cultures. J. Vis. Exp. (45), e2384, doi:10.3791/2384 (2010).
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