We would like to establish an in vitro chronic, long-lasting ischemia model in primary (mouse or rat) neuronal cultures with CoCl2. Literatures suggest 1 uM CoCl2 for 7 days detain OPC (oligodendrocyte precursor cell) maturation, and 200-2000 uM CoCl2 leads to 30-50% neuronal death in 1-4 hours. Does anyone have any experience or suggestion on long term (i.e. days) CoCl2 treatment in primary neurons, such as the CoCl2 concentration or duration? Thanks!
Hello Tuo Yang.
It will depend on the aims of the experiments. Are you interested in cell phenotype caused by ishemic condition or time-course ROS accumulation and subsequent cell proliferation activity or cell death or apoptotic activation of genes related to stress, inflammatory responses? Or would you like to rescue the ischemic cells by adding anti-ischemic agents? Depending on the purposes, you may need to select lower concentration for 7 days as an end point or 24~48h at higher concentration as endpoints.
To do perfect expts in which you could avoid the abrupt toxic effects of the CoCl2, obtain a concentration of IC50 or LD50 by MTT assay or propidium iodide and Hoechst dye, respectively, for your specific cells in YOUR hands,
Best wishes.
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