Treatment for Chemotherapy-Induced Peripheral Neuropathy
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[0032]Neuronal Cell Culture. Mixed cortical and striatal neurons fromembryonic day 17 male Sprague-Dawley rats (tissue obtained from NIH) were seeded into poly-D-lysine coated 96-well plates at density of 50,000 cells / well and initially cultured at 37° C., 5% CO2, in neurobasal medium (NB) with B27 supplement (Invitrogen) containing full strength of AOs to obtain highly enriched (95%) neurons. Since AOs, including vitamin E, vitamin E acetate, superoxide dismutase (SOD), catalase (CAT), and GSH, are additives to culture medium, reduction of AO concentration in culture medium provides an approach to determine the level of OS involvement in a neurotoxic process. In our study, the culture medium was replaced at a 50% ratio with NB plus B27 minus AOs twice at days 7 and 9 to set AO concentrations as 50% and 25%, respectively. At 16 days in vitro (d.i.v.), a fraction of the culture medium was harvested for lactate dehydrogenase (LDH) assay, and then AO concentration ...
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