Live imaging of primary cerebral cortex cells using a 2D culture system

Bruna Soares Landeira, Jéssica Alves de Medeiros Araújo, Timm Schroeder, Ulrich Müller, Marcos R. Costa

Research output: Contribution to journalArticlepeer-review

Abstract

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. Video Link.

Original languageEnglish (US)
Article numbere56063
JournalJournal of Visualized Experiments
Volume2017
Issue number126
DOIs
StatePublished - Aug 1 2017

Keywords

  • Cell proliferation
  • Cell survival
  • Cerebral cortex
  • Issue 126
  • Neurobiology
  • Neuronal differentiation
  • Primary cell culture
  • Time-lapse video-microscopy

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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