There are two main methods for preparing the PuraMatrix (PM) as a three-dimensional (3-D) scaffold for supporting the cells including surface plating and encapsulation. In the surface-plating procedure, the scaffold is created by adding an ionic agent such as medium to PM. After adjusting the PH by changing the medium, cells are seeded on top of the scaffold. In the encapsulation method, PM and cells are mixed suddenly to produce a true 3-D culture.  Because PM is very acidic agent, it can be harmful for sensitive cells such as neural stem cells. We introduced a new method for solving this problem. In this novel approach, firstly, scaffold was created, then, the neural stem cells were injected into the several sites of it. In this method, the survival of the cells was significantly high than that of other methods. This method can be used in future investigations in which a 3-D culture of neural stem cells in needed.