Neuronal pc12 rat adrenal pheochromocytoma cell line

A mouse macrophage cell line (Raw 264.7) was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA, cat # TIB-71), and cultured in Dulbecco’s Modified Eagle’s Media (DMEM) (Invitrogen) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS). Neuronal PC12 rat adrenal pheochromocytoma cell line was obtained from ATCC, and cultured in Dulbecco’s modified Eagle medium (Hyclone, South Logan, UT, USA) supplemented with 10 % FBS, and 1 % (v/v) of both penicillin and streptomycin. The cells were grown in an incubator with optimal culture conditions of 37 °C and 5% CO₂, and the medium was routinely replaced every 2–3 days.
Bone marrow derived macrophages (BMM) were obtained by differentiation of bone marrow stem cells extracted from murine femurs (C57BL/6, female mice) as described in [56 (link)]. The cells were cultured for 10 days in media supplemented with 1000 U/mL macrophage colony-stimulating factor (MCSF). The purity of monocyte culture was determined by flow cytometry using FACS Calibur (BD Biosciences, San Jose, CA, USA). Mouse primary cultured cortical neurons and dopaminergic (DA) neurons from substantia nigra pars compacta (SNpc) were isolated from mouse pups cortex and midbrain as described [57 ].