Powder bed fusion using a laser beam (PBF-LB) offers a suitable alternative to manufacturing Ti65 with intricate geometries and internal structures in hypersonic aerospace applications. However, issues such as undesirable surface roughness, defect formation, and microstructural inhomogeneity remain critical barriers to its wide application. In this study, a coupled discrete element method–computational fluid dynamics (DEM-CFD) model was utilized to investigate the spreading behavior of Ti65 powder in a multi-layer PBF-LB process. The macro- and microscopic characteristics of the powder beds were systematically analyzed across different layers and regions under various spreading velocities. The results show that the packing density and uniformity of the powder beds in multi-layer PBF-LB of Ti65 powder improves as the number of solidified layers increases. Poor bed quality is observed in the first two layers due to a strong boundary effect, while a stable and denser powder bed emerges from the fourth layer. The presence of a previously solidified region strongly influences its neighboring unsolidified areas, enhancing density in the upstream region and causing looser packing downstream. Additionally, due to the existence of a solidified region, the height of the powder bed progressively decreases along the spreading direction.