The combination of cell microenvironment control and real-time monitoring of cell signaling events can provide key biological information. Through precise multipatterning of gold nanoparticles (GNPs) around cells, sensing and actuating elements can be introduced in the cells' microenviroment, providing a powerful substrate for cell studies. In this work, a combination of techniques are implemented to engineer complex substrates for cell studies. Alternating GNPs and bioactive areas are created with micrometer separation by means of a combination of vacumm soft-lithography of GNPs and protein microcontract printing. Instead of conventional microfluidics that need syringe pumps to flow liquid in the microchannels, degas driven flow is used to fill dead-end channels with GNP solutions, rendering the fabrication process straightforward and accessible. This new combined technique is called Printing and Vacuum lithography (PnV lithography). By using different GNPs with various organic coating ligands, different macroscale patterns are obtained, such as wires, supercrystals, and uniformly spread nanoparticle layers that can find different applications depending on the need of the user. The application of the system is tested to pattern a range of mammalian cell lines and obtain readouts on cell viability, cell morphology, and the presence of cell adhesive proteins.