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A Cell Culture Model for Studying the Role of Neuron-Glia Interactions in Ischemia.

Authors
  • Gava-Junior, Genilso1
  • Roque, Cláudio1
  • Mendes-Oliveira, Julieta1
  • Bernardino, Ana C1
  • Serrenho, Inês1
  • Pires, Joel P1
  • Baltazar, Graça2
  • 1 Centro de Investigação em Ciências da Saúde (CICS-UBI), Universidade da Beira Interior.
  • 2 Centro de Investigação em Ciências da Saúde (CICS-UBI), Universidade da Beira Interior; Faculdade de Ciências da Saúde, Universidade da Beira Interior; [email protected]
Type
Published Article
Journal
Journal of Visualized Experiments
Publisher
MyJoVE Corporation
Publication Date
Nov 14, 2020
Issue
165
Identifiers
DOI: 10.3791/61388
PMID: 33252110
Source
Medline
Language
English
License
Unknown

Abstract

Ischemic stroke is a clinical condition characterized by hypoperfusion of brain tissue, leading to oxygen and glucose deprivation, and the consequent neuronal loss. Numerous evidence suggests that the interaction between glial and neuronal cells exert beneficial effects after an ischemic event. Therefore, to explore potential protective mechanisms, it is important to develop models that allow studying neuron-glia interactions in an ischemic environment. Herein we present a simple approach to isolate astrocytes and neurons from the rat embryonic cortex, and that by using specific culture media, allows the establishment of neuron- or astrocyte-enriched cultures or neuron-glia cultures with high yield and reproducibility. To study the crosstalk between astrocytes and neurons, we propose an approach based on a co-culture system in which neurons cultured in coverslips are maintained in contact with a monolayer of astrocytes plated in multiwell plates. The two cultures are maintained apart by small paraffin spheres. This approach allows the independent manipulation and the application of specific treatments to each cell type, which represents an advantage in many studies. To simulate what occurs during an ischemic stroke, the cultures are subjected to an oxygen and glucose deprivation protocol. This protocol represents a useful tool to study the role of neuron-glia interactions in ischemic stroke.

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