Introduction: Recent studies have shown the key roles of different sequences and motifs in the extracellular matrix guidance. Biological activities of the macromolecules can be mimicked by the motifs of extracellular matrix. Laminin peptide sequences and motifs can bind the biomaterials of scaffolds, improve the cells mechanical protection, increase the cells growth, and lead to the formation of tissues. Materials and Methods: Neural stem cells (NSCs) were isolated from embryonic ganglionic eminences of 14 days rats and cultured as neurosphere. The number and diameter of neurospheres on days 3, 7, and 10 were measured. NSCs were cultured by two-dimensional (2D) and 3D cultures. To evaluate cell survival, Live/Dead kit was used on days 1 and 7. The cell proliferation was assessed using the MTS assay on days 1 and 7. Results: Our finding showed that the cells isolated from ganglionic eminences have a high proliferative capacity. According to the proliferation assay measurements, the number and diameter of neurosphere significantly increased after culturing for 10 days. Cell proliferation and survival in the self-assembling peptide nanofiber containing laminin motif (SAPN-LM) were significantly higher than that of 2D. Conclusion: SAPN-LM improved cell proliferation and survival and suggest an appropriate models for further investigations.