Hi all.
I'm planning to incubate my primary hippocampal neuron culture in microglia-conditioned media, in which the microglia were stimulated with LPS+ATP to produce cytokines. As we use different media for each culture (Neurobasal medium supplemented with B-27, 0.5% fetal bovine serum, 0.5 mm Glutamax and sodium pyruvate for rat primary neuron culture; DMEM/F12 supplemented with 10% FBS and glutamax for mouse mixed glia culture, from which microglia were isolated by trysinization and maintained in mixed glia-conditioned media afterward), I want to know which culture media to use for this media-transfer experiments. (stimulate microglia with LPS in neurobasal media vs DEME/F12? media or else?)
I'm not really familiar with glia culture and experiments using conditioned media, so I'm looking forward some advice. I would be glad if anyone suggest me some reference paper or protocol. Any comments will be appreciated.
Thanks in advance👍