Cultured PC12 cells: A model for neuronal function, differentiation, and survival

This chapter presents a model for neuronal function, differentiation, and survival of cultured pheochromocytoma PC12 cell lines. The optimal final dilution of the collagen should be determined empirically by testing various concentrations for their capacities to foster cell attachment and nerve growth factor (NGF)-promoted neurite outgrowth. Spread the collagen evenly over the surface of the culture dish with the use of an L-shaped glass rod. Dry collagen by leaving the plates uncovered for 1-2 h in a tissue culture hood. One needs to store collagen-coated dishes at room temperature and use within one week after preparation. Dislodge the cells from the surface of the dish by repeated and forceful discharge of the culture medium directly onto the cells with a disposable glass Pasteur pipette. Forceful trituration of the cell suspension within the Pasteur pipette also decreases cell clumping. To determine the numbers of PC12 cells suspended in culture or other medium, pellet the cells by centrifugation, aspirate to remove the medium, and resuspend the cells in a known volume of the working nuclei-counting solution.