The invention provides a preparation method of neural stem cells and glutamatergic neurons in cerebral cortex, and relates to an iPSC complete neural induction culture medium. The iPSC complete neuralinduction culture medium comprises the following components: a cell basic culture medium, a cell culture additive, neural development factors and an inhibitor. The invention provides the iPSC complete neural induction culture medium, an iPSC nerve cell culture medium and a culture method. The culture medium includes a serum-free basic medium, a cell additive, neural development factors and an inhibitor. The use of the serum-free basic medium can avoid the introduction of exogenous substances and reduce the risk of pollution. Meanwhile, a variety of nutrients, such as the cell additive and theneural development factors, can enable iPSC cells to be stimulated by signals so that iPSC cells can be activated to promote differentiation. The cell culture method provided by the invention is simple, efficient, low in cost, and can induce high-quality neuron cells in a short period.