An Ex Vivo Technique to Induce Neuronal Death via Oligodendrocyte-Specific CD8+ T Cells

Published: August 31, 2023

Abstract

Source: Göbel, K., et al. An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices. J. Vis. Exp. (2015).

This video demonstrates an ex vivo technique to induce brain cell death mediated through cytotoxic CD8+ T cells. Upon obtaining brain slices, activated oligodendrocyte-directed CD8+ T cells are transferred onto the slices to target the myelin sheath and oligodendrocytes. Post-incubation, the damage to myelin and neuronal death can be assessed histologically in the regions of interest.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. General Comments for Mouse Experiments Keep the mice under pathogen-free conditions and enable them access to food and water ad libitum. NOTE: It is important to use age- and sex-matched mice in experimental groups because immunological patterns can vary with age and gender. <…

Disclosures

The authors have nothing to disclose.

Materials

12-Well-plate Corning 3513
2-Mercaptoethanol Gibco 31350-010
2-Methylbutan Roth 3927.1
70 µm strainer Falcon 352350
CaCl2 Merck 1.02382.0500 Calcium chloride
CD8+-isolation kit Miltenyi Biotech 130-090-859
D(+)-glucose Merck 1.08337.1000
DMEM Gibco 31966-021 Warm in 37 °C water bath before use
EDTA Sigma E5134
FCS PAA Laboratories A15-151 Fetal calve serum
Gentamicin Gibco 15750-060
HEPES 1M Gibco 15630-050
IL-2 Peprotech 212-12
Isofluran Abbott 05260-05
KCl Merck 1.04933.0500 Potassium chloride
KHCO3 Sigma P9144 Potassium hydrogen carbonate
L-Glutamine Gibco 35050-038
MgSO4 Merck 1.05886.0500 Magnesium sulfate
NaCl Sigma 31434 Sodium chloride
NaH2PO4 * H2O Merck 1.06346.0500 Sodium hydrogen phosphate
NaHCO3 Merck 1.06329.0500 Sodium hydrogen carbonate
NaOH Merck 1.09137.1000 Sodium hydroxide
NH4Cl Sigma 213330 Ammonium chloride
Non-Essential Amino Acid Gibco 11140-050
OVA (257-264) Genscript RP10611 Ovalbumin
PIPES Sigma P6757
Sukrose Merck 1.07687.1000
Tissue-Tek OCT Sakura 4583

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Cite This Article
An Ex Vivo Technique to Induce Neuronal Death via Oligodendrocyte-Specific CD8+ T Cells. J. Vis. Exp. (Pending Publication), e21608, doi: (2023).

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