Article Dans Une Revue Journal of visualized experiments : JoVE Année : 2020

Generation of Oligodendrocytes and Oligodendrocyte-Conditioned Medium for Co-Culture Experiments

Résumé

In the central nervous system, oligodendrocytes are well-known for their role in axon myelination, that accelerates the propagation of action potentials through saltatory conduction. Moreover, an increasing number of reports suggest that oligodendrocytes interact with neurons beyond myelination, notably through the secretion of soluble factors. Here, we present a detailed protocol allowing purification of oligodendroglial lineage cells from glial cell cultures also containing astrocytes and microglial cells. The method relies on overnight shaking at 37 °C, which allows selective detachment of the overlying oligodendroglial cells and microglial cells, and the elimination of microglia by differential adhesion. We then describe the culture of oligodendrocytes and production of oligodendrocyte-conditioned medium (OCM). We also provide the kinetics of OCM treatment or oligodendrocytes addition to purified hippocampal neurons in co-culture experiments, studying oligodendrocyte-neuron interactions.

Fichier principal
Vignette du fichier
Mazuir JoVE.pdf (189.44 Ko) Télécharger le fichier
Origine Fichiers produits par l'(les) auteur(s)
Licence

Dates et versions

hal-03052042 , version 1 (12-01-2021)

Licence

Identifiants

Citer

Elisa Mazuir, Anne-Laure Dubessy, Loane Wallon, Marie-Stéphane Aigrot, Catherine Lubetzki, et al.. Generation of Oligodendrocytes and Oligodendrocyte-Conditioned Medium for Co-Culture Experiments. Journal of visualized experiments : JoVE, 2020, 156, pp.e60912. ⟨10.3791/60912⟩. ⟨hal-03052042⟩
156 Consultations
827 Téléchargements

Altmetric

Partager

  • More