Skip to Content
Merck
  • Methods for culturing adult CNS neurons reveal a CNS conditioning effect.

Methods for culturing adult CNS neurons reveal a CNS conditioning effect.

Cell reports methods (2022-07-27)
Erna A van Niekerk, Riki Kawaguchi, Camila Marques de Freria, Kimberly Groeniger, Maria C Marchetto, Sebastian Dupraz, Frank Bradke, Daniel H Geschwind, Fred H Gage, Mark H Tuszynski
ABSTRACT

Neuronal cultures provide a basis for reductionist insights that rely on molecular and pharmacological manipulation. However, the inability to culture mature adult CNS neurons limits our understanding of adult neuronal physiology. Here, we report methods for culturing adult central nervous system neurons in large numbers and across multiple brain regions for extended time periods. Primary adult neuronal cultures develop polarity; they establish segregated dendritic and axonal compartments, maintain resting membrane potentials, exhibit spontaneous and evoked electrical activity, and form neural networks. Cultured adult neurons isolated from different brain regions such as the hippocampus, cortex, brainstem, and cerebellum exhibit distinct cell morphologies, growth patterns, and spontaneous firing characteristics reflective of their regions of origin. Using adult motor cortex cultures, we identify a CNS "conditioning" effect after spinal cord injury. The ability to culture adult neurons offers a valuable tool for studying basic and therapeutic science of the brain.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Fezf2 Antibody, from rabbit, purified by affinity chromatography
Sigma-Aldrich
Anti-Tau-1 Antibody, clone PC1C6, clone PC1C6, Chemicon®, from mouse
Sigma-Aldrich
Laminin from Engelbreth-Holm-Swarm murine sarcoma basement membrane, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Anti-Vesicular Glutamate Transporter 1 Antibody, serum, Chemicon®
Sigma-Aldrich
Anti-Choline Acetyltransferase Antibody, Chemicon®, from goat