With its high mechanical strength and its remarkable thermal and electrical properties, suspended graphene has long been expected to find revolutionary applications in optoelectronics or as a membrane in nano-devices. However, the lack of efficient transfer and patterning processes still limits its potential. In this work, we report an optimized anthracene-based transfer process to suspend few layers of graphene (1-, 2- and 4-layers) in the millimeter range (up to 3 mm) with high reproducibility. We have explored the advantages and limitations for patterning of these membranes with micrometer-resolution by focused ion beam (FIB) and picosecond pulsed laser ablation techniques. The FIB approach offers higher patterning resolution but suffers from the low throughput. We demonstrate that cold laser ablation is a fast and flexible method for micro-structuring of suspended graphene. One promising field of application of ultimately thin, microporous graphene membranes is their use as next-generation cell culture supports as alternative to track-etched polymer membranes, which often exhibit poor permeability and limited cell-to-cell communication across the membranes. To this end, we confirmed good adhesion and high viability of placental trophoblast cells cultivated on suspended porous graphene membranes without rupturing of the membranes. Overall, there is high potential for the further development of ultrathin suspended graphene membranes for many future applications, including their use for biobarrier cell culture models to enable predictive transport and toxicity assessment of drugs, environmental pollutants, and nanoparticles.