Prenatal cerebrospinal fluid modulate differentiation and proliferation of rat pheochoromocytoma PC12 cells

Background During the early stages of brain development, ependymal cells that line the neural tube are thought to secrete CSF. It is well documented that fetal CSF contains many neurotrophic and growth factors which are known as modulators of neurogenesis, differentiation and brain extracellular microenvironment. Rat pheochromocytoma PC12 cells have been widely used as an in vitro model of neuronal differentiation since the cells undergo differentiation to sympathetic neuron-like cells in response to NGF, bFGF, EGF, TGF-a and GDNF. We hypothesized that prenatal CSF could have differentiational effect on PC12 cells. Materials and methods CSF was removed by tapping the cisterna magna of Wistar rat fetuses (E17- E20) then centrifuged at 4000 r.p.m for 10 min, the supernatant frozen immediately and stored at -70°C until used. PC12 cells were cultured in RPMI-1640 with 10% FBS, 100 unit/ml of penicillin, 100 mg/ml of streptomycin and 5% CO2 at 37°C. For culture experiments, 2*10E4 PC12 cells were added to each well of a 96-well plate that had been coated with Poly-D-Lysine. After attachment, the cells were exposed to CSF at different ages with dissimilar concentration of 7, 10, 25% (v/v). The cell viability and cell proliferation were measured by MTT assay. The neuronal differentiation of PC12 cells were considered by changes of neurite outgrowth. Results