I-BOP

Primary embryonic hippocampal neuron culture is an in vitro method in which hippocampi from embryonic rodents are dissected, enzymatically or mechanically dissociated, plated on adhesive substrates, and maintained in defined neuronal medium or in low-density sandwich/co-culture formats to support neuronal attachment, neurite extension, polarity formation, dendritic arborization, and synapse formation. The main readouts are cell survival, neuronal purity, neurite outgrowth, axon-dendrite polarization, synaptic marker development, and functional neuronal activity, assessed by phase-contrast microscopy, immunocytochemistry for neuronal/glial markers, live imaging, or electrophysiology depending on the downstream experiment.

The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.