Generation of Neurospheres from Mixed Primary Hippocampal and Cortical Neurons Isolated from E14-E16 Sprague Dawley Rat Embryo
Primary Neuron Culture is an essential technique in the field of neuroscience. In order to gain deeper mechanistic insights of the brain, it is essential to have a robust in vitro model which could be exploited for various neurobiology studies. Though primary neurons cultures like long term hippocampal cultures have provided scientists with a model, yet it does not represent the complexity of brain network completely. In the wake of these limitations of the primary neuron culture, a new model called neurospheres bearing a closer resemblance to the brain tissue has emerged. In the present protocol, just by plating high and low densities of mixed cortical and hippocampal neurons isolated from the embryo of an E14-E16 (Embryonic Day 14-16) Sprague Dawley Rat, we were able to generate neurospheres and long-term primary neuron culture as two independent platforms to conduct further studies. This process is extremely simple and cost effective as it does away with several steps and reagents previously deemed essential for the neuron culture. This is a robust protocol with minimum requirements that can be performed with easily achievable results and can be used for a diversity of studies related to neuroscience.