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Culture of primary cortical and hippocampal neurons

AT Alejandro López Tobón
MS Megalakshmi Suresh
JJ Jing Jin
AV Alessandro Vitriolo
TP Thorben Pietralla
KT Kerry Tedford
MB Michael Bossenz
KM Kristina Mahnken
FK Friedemann Kiefer
GT Giuseppe Testa
KF Klaus-Dieter Fischer
AP Andreas W. Püschel
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Dissociated cortical and hippocampal neurons from embryonic day 18 (E18) rat or E17 mouse embryos were prepared and transfected by calcium phosphate co-precipitation as described previously5,71. Neurons were plated at 70,000 cells per well in a 24-well plate coated with poly-L ornithine in Neurobasal medium (Life Technologies) and cultured at 37 °C and 5% CO2. The culture medium was replaced by 400 μl Opti-MEM (Life Technologies) before the DNA/ CaCl2 mixture was added. After incubation for 45 min at 37 °C and 5% CO2 the neurons were washed for 15 min with 1 ml opti-MEM, which had been pre-incubated at 37 °C and 10% CO2, and Neurobasal medium was added back to the cells.

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