Abstract: The Stellite6 Co-based alloy layers were fabricated on 316L stainless steel substrate using laser wire feeding cladding. The formation, dilution rate, microstructure, chemical composition, microhardness and wear resistance of the cladding layers were analyzed. The results indicate that the two-layer laser cladding process can ensure the optimal formation of the cladding layer with a dilution rate below 5%, and the dilution rate is uniform. The microstructure of the cladding layer is a subeutectic structure composed of dendritic Co-rich solid solution and network eutectic carbides, and it has a higher concentration of network carbides at the top layer compared to the bottom layer. The two-layer cladding process results in a stepwise increase of hardness in the vertical direction of the cladding layer, which stabilizes after reaching a thickness of 1.8mm. The horizontal microhardness of the cladding layer near the surface is relatively uniform, ranging from 450 HV to 550HV. The two-layer cladding process demonstrates superior wear resistance, with a friction coefficient of 0.37 and a 53.9%reduction of mass loss compared to the base material. The wear mechanism of the two-layer cladding process changes from adhesive wear of the substrate to abrasive wear.