Pulsed Nd:YAG laser was used to microtexture AISI-O1 cold work steel to control the substrate surface characteristics for later thermal spray coating to enhance the wearability. Influence of microtexturing parameters on the 3D surface morphology was studied by Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM), and Optical Microscopy (OM). Results show that when AISI-O1 specimens are irradiated with various parameters the morphology of AISI-O1 cold work steel is changed correspondingly. It demonstrates that for a given laser the influence of laser microtexturing parameters on the morphology of the machined surface could be established successfully attractive to the thermal spray world. In the meantime, the wearability and antifatigue of the succedent thermal spray coating are significantly increased with the appropriate microtextured surface.