The effects of fetal rat cerebrospinal fluid (CSF) on neural differentiation of rat pheochoromocytoma PC12 cells

Fetal cerebro- spinal fluid (CSF) contains many neurotrophic and growth factors. Rat pheochromocytoma PC12 cells have been widely used as an in vitro model of neuronal differentiation that undergo differentiation to sympathetic neuron-like cells in response to NGF, bFGF, EGF and GDNF. CSF was removed by tapping the cisterna magna of Wistar rat fetuses (E17- E20). PC12 cells were cultured in RPMI plus 10% FBS. The cell viability and cell proliferation were measured by MTT assay. Neural differentiation markers (MAP-2 and β-III tubulin) expressions were analyzed by immunocytochemistry. MAP-2 and β-III tuobulin were expressed in PC12 cells cultured in CSF supplemented medium, but not in the cells from control cultures. Viability and cell proliferation were significantly elevated in PC12 cells cultured in CSF supplemented medium in E18 compared with control ones. A significant neuronal-like outgrowth appeared as early as Day 3 after the application of the CSF supplemented medium E17 and E19. Our data are in the same line with pervious studies that clarify crucial role of CSF neurotrophic factors in neuronal differentiation. Taken together we address PC12 neuronal differentiation to CSF induction by its components especially growth factors.