Culturing Newborn DRG Neurons

DRGs were obtained from newborn (postnatal days 0–2) WT and Gdap1 knockout (Gdap1^−/−) mice according to (45 (link)). Ganglia were digested in collagenase (Worthington, USA) and trypsin (Sigma-Aldrich, St. Louis, MO), dissociated by trituration and plated in DMEM-F12 containing 50 ng/ml NGF (Tebu-Bio, Le-Perray-en-Yvelines, France), 10% FBS and 5 ng/ml of aphidicolin (A.G. Scientific, San Diego, CA). DRG neurons were seeded on poly-l-lysine and laminin (1 μg/ml)-coated (Sigma-Aldrich, St. Louis, MO) plates and used for experimentation between 2 and 4 DIV.

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Nuevo-Tapioles C., Santacatterina F., Sánchez-Garrido B., de Arenas C.N., Robledo-Bérgamo A., Martínez-Valero P., Cantarero L., Pardo B., Hoenicka J., Murphy M.P., Satrústegui J., Palau F, & Cuezva J.M. (2021). Effective therapeutic strategies in a preclinical mouse model of Charcot–Marie–Tooth disease. Human Molecular Genetics, 30(24), 2441-2455.

Publication 2021

collagenase trypsin aphidicolin poly-l-lysine laminin

Aphidicolin Collagenase Ganglia Laminin Lysine Mice Neurons Newborn Poly Trypsin

Corresponding Organization : Consejo Superior de Investigaciones Científicas

Other organizations : Hospital Sant Joan de Déu Barcelona, MRC Mitochondrial Biology Unit, Medical Research Council, University of Cambridge, Wellcome Trust, Universitat de Barcelona, Hospital Clínic de Barcelona

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Variable analysis

independent variables

Genotype (WT vs. Gdap1 knockout)

dependent variables

Not explicitly mentioned

control variables

Postnatal age (0-2 days)
Culture conditions (DMEM-F12 with 50 ng/ml NGF, 10% FBS, 5 ng/ml aphidicolin)
Substrate (poly-L-lysine and laminin-coated plates)
Time in culture (2-4 DIV)

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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