Murine DRG explants dissected from embryonic day (E) 13 C56BL/6J mice were cultured on poly-L-lysine (PLL) and laminin-coated 24-well plates in Neuro-medium (Miltenyi Biotec) containing 10% FBS and 25 ng/ml nerve growth factor (NGF) (Harlan Bioproducts). After 24 h, the culture medium was changed to Neuro-medium supplemented with 2% Neuro-Brew-21 (Miltenyi Biotec), 25 ng/ml NGF, and 1 mM Glutamine in addition to a mixture of 1 mM 5′-fluoro-2′-deoxyuridine and 1 mM uridine to remove non-neuronal cells. The in vitro Wallerian degeneration of axons was introduced by removing cell bodies at 10–14 d in vitro using a pipette tip.5,6 (link)
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Murine DRG Explant Culture for Axon Degeneration
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Axons
Cell bodies
Deoxyuridine
Embryonic
Glutamine
Laminin
Lysine
Mice
Nerve growth factor
Non neuronal cells
Poly
Uridine
Wallerian degeneration
Corresponding Organization : National Center of Neurology and Psychiatry
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Variable analysis
independent variables
- Removing cell bodies at 10–14 d in vitro using a pipette tip
- Wallerian degeneration of axons
- Culture medium with Neuro-medium, 2% Neuro-Brew-21, 25 ng/ml NGF, and 1 mM Glutamine
- Mixture of 1 mM 5′-fluoro-2′-deoxyuridine and 1 mM uridine to remove non-neuronal cells
- Culturing murine DRG explants on poly-L-lysine (PLL) and laminin-coated 24-well plates
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