CSIR-Indian Institute of Chemical Biology 1 article published in JoVE Neuroscience Generation of Neurospheres from Mixed Primary Hippocampal and Cortical Neurons Isolated from E14-E16 Sprague Dawley Rat Embryo Gaurav Das1,2, Varsha Gupta1, Juhee Khan1, Deepshikha Mukherjee1, Surajit Ghosh1,2 1Organic and Medicinal Chemistry Division, CSIR-Indian Institute of Chemical Biology, 2Academy of Scientific and Innovative Research (AcSIR) Presented here is a protocol for the spontaneous generation of neurospheres enriched in neural progenitor cells from high density plated neurons. During the same experiment, when neurons are plated at a lower density, the protocol also results in prolonged primary rat neuron cultures.