Dissociated cortical neurons were prepared from embryonic day 17-18 (E17-18) rat cerebral cortex as described previously37 (link),51 (link),52 (link) and cultured in Neurobasal medium (Life Technologies, Carlsbad, CA) supplemented with B27 (Life Technologies), glutamine (Sigma, St. Louis, MO) and penicillin-streptomycin (Sigma). Neurons were plated on poly-D-lysine (BD Biosciences, San Jose, CA) and laminin (BD Biosciences) coated glass coverslips (12 mm, #1.5; Cat#: 64-0712, Warner Instruments, Camden, CT) or in glass bottom 35mm dishes made with #1.5 German optic cover glass (cat#: GBD00004-200, Cell E&G LLC, Houston, TX). Neurons were plated at 150,000/well in 24-well plates or at 180,000 neurons per 18 mm glass coverslip of a single 35 mm dish for transfection experiments and were maintained in a humidified 37°C incubator with 5% CO2.
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Dissociated Cortical Neuron Culture Protocol
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Cell
Cerebral cortex
Cultured medium
Dish
Embryonic
Glutamine
Laminin
Lysine
Neurons
Optic
Penicillin
Poly
Streptomycin
Transfection
Corresponding Organization :
Other organizations : Thomas Jefferson University, University of Delaware, Imaging Center
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Variable analysis
independent variables
- Embryonic day 17-18 (E17-18) rat cerebral cortex
- Not explicitly mentioned
- Neurobasal medium (Life Technologies, Carlsbad, CA) supplemented with B27 (Life Technologies), glutamine (Sigma, St. Louis, MO) and penicillin-streptomycin (Sigma)
- Poly-D-lysine (BD Biosciences, San Jose, CA) and laminin (BD Biosciences) coated glass coverslips (12 mm, #1.5; Cat#: 64-0712, Warner Instruments, Camden, CT) or in glass bottom 35mm dishes made with #1.5 German optic cover glass (cat#: GBD00004-200, Cell E&G LLC, Houston, TX)
- Plating density (150,000/well in 24-well plates or at 180,000 neurons per 18 mm glass coverslip of a single 35 mm dish)
- Humidified 37°C incubator with 5% CO2
- No positive or negative controls were explicitly mentioned in the protocol.
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