Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System

Landeira, Bruna Soares; Araújo, Jéssica Alves de Medeiros; Schroeder, Timm; Müller, Ulrich; Costa, Marcos Romualdo

Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System

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dc.contributor.author	Landeira, Bruna Soares
dc.contributor.author	Araújo, Jéssica Alves de Medeiros
dc.contributor.author	Schroeder, Timm
dc.contributor.author	Müller, Ulrich
dc.contributor.author	Costa, Marcos Romualdo
dc.date.accessioned	2017-08-10T13:19:13Z
dc.date.issued	2017-08-09
dc.description.embargo	2019-08-09
dc.description.resumo	During cerebral cortex development, progenitor cells undergo several rounds of symmetric and asymmetric cell divisions to generate new progenitors or postmitotic neurons. Later, some progenitors switch to a gliogenic fate, adding to the astrocyte and oligodendrocyte populations. Using time-lapse video-microscopy of primary cerebral cortex cell cultures, it is possible to study the cellular and molecular mechanisms controlling the mode of cell division and cell cycle parameters of progenitor cells. Similarly, the fate of postmitotic cells can be examined using cell-specific fluorescent reporter proteins or post-imaging immunocytochemistry. More importantly, all these features can be analyzed at the single-cell level, allowing the identification of progenitors committed to the generation of specific cell types. Manipulation of gene expression can also be performed using viral-mediated transfection, allowing the study of cell-autonomous and non-cell-autonomous phenomena. Finally, the use of fusion fluorescent proteins allows the study of symmetric and asymmetric distribution of selected proteins during division and the correlation with daughter cells fate. Here, we describe the time-lapse video-microscopy method to image primary cerebral cortex murine cells for up to several days and analyze the mode of cell division, cell cycle length and fate of newly generated cells. We also describe a simple method to transfect progenitor cells, which can be applied to manipulate genes of interest or simply label cells with reporter proteins.	pt_BR
dc.identifier.doi	10.3791/56063
dc.identifier.uri	https://repositorio.ufrn.br/jspui/handle/123456789/23745
dc.language	eng	pt_BR
dc.rights	Acesso Restrito	pt_BR
dc.subject	Neurobiology	pt_BR
dc.subject	Primary cell culture	pt_BR
dc.subject	cerebral cortex	pt_BR
dc.subject	time-lapse video-microscopy	pt_BR
dc.subject	cell proliferation	pt_BR
dc.subject	neuronal differentiation	pt_BR
dc.subject	cell survival	pt_BR
dc.title	Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System	pt_BR
dc.type	article	pt_BR