Culturing Murine Microglia and Human Neuroblastoma Cells

The murine microglial line BV2 was a generous gift from Prof. E. Blasi (Università degli Studi di Modena e Reggio Emilia, Italy); the human neuroblastoma SH-SY5Y line was purchased from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK). Both lines were cultured in DMEM medium supplemented with 5% fetal calf serum and 100 μM nonessential amino acids, 2 mM L-alanyl-L-glutamine, and 50 mg/ml penicillin-streptomycin (all from Thermo Fisher Scientific, UK) at 37°C in a 5% CO₂ atmosphere. SH-SY5Y cells were differentiated to a neuron-like phenotype prior to experimentation by incubation with 10 μM trans-retinoic acid (Sigma-Aldrich, UK) for 5 days [26 (link)].
Primary murine microglia were prepared from C57Bl/6 male mice aged 8 weeks according to our previously published protocols [27 (link)]. Cells were cultured in DMEM medium supplemented with 20% fetal calf serum and 100 μM nonessential amino acids, 2 mM L-alanyl-L-glutamine, and 50 mg/ml penicillin-streptomycin (all Thermo Fisher Scientific, UK) at 37°C in a 5% CO₂ atmosphere.