A Simplified Method for Ultra-Low Density, Long-Term Primary Hippocampal Neuron Culture

The overall goal of this procedure is to establish healthy longterm cultures of primary mouse hippocampal neurons at ultra-low density, in order to facilitate immunocytochemistry labeling, and the study of neuronal cell autonomous mechanisms. This study can help understand some key questions in the field of neuroscience research such as the studies on the specificity of protein in neuromorphology of function of development. The main advantage of this technique is that it allows neurons to grow at ultra-low density on a cover slip without the support of a glial feeder layer.

Demonstrating the procedure will be Mariel Piechowicz, a student from my laboratory. Prepare two high-density and two low-density 24-well plates. Using a 28 gauge needle, etch the bottom of 24 well plates, so that the displaced plastics will act as a support for the cover slips with low-density neurons growing on them.