Molecular patterning on biomaterial surfaces is an effective strategy to regulate biomaterial properties. Among the specific molecules, due to their biological functions, such as regulating cell behavior, heparin-like polymers (HLPs) have attracted much attention. In this study, HLP-distributed regional patterned surfaces (300 μm diameter circular array) were prepared by the combination of visible light-induced graft polymerization, transfer imprinting, and self-assembly to regulate the behavior of human umbilical vein endothelial cells (HUVECs) and human umbilical vein smooth muscle cells (HUVSMCs). The introduction of the regional pattern on HLP-modified surfaces enhanced the promotion effect of sulfonate-containing polymer (pSS) and sulfonate-, and glyco-containing copolymer (pS-co-pM), and slightly weakened the inhibition effect of glyco-containing polymer (pMAG) on the growth of HUVECs and HUVSMCs. Compared with flat surfaces, it was found that the unmodified regional patterned surfaces inhibit the spreading of HUVECs and HUVSMCs, while significantly promoting the spreading of HUVECs and HUVSMCs on all the HLP-distributed regional patterned surfaces. The patterned surface modified with pS-co-pM had the highest average spread area of HUVECs (∼10,554 μm2), which was 193 % higher than that of the unmodified flat surface. This trend was somewhat related to surface VEGF adsorption. The combination of regional divisive patterns and different HLP distributions enriched the potential of further exploring the influences of HLP chemical distributions and complex surface environments on cell-material interactions.