Cellular model systems are essential platforms used across multiple research fields for exploring the fundaments of biology and biochemistry. Here, we present giant plasma membrane vesicles (GPMVs) as a platform of cell-like compartments that will facilitate the study of particles within a biorelevant environment and promote their further development. We studied how cellularly taken up nanoparticles (NPs) can be transferred into formed GPMVs and which are the molecular factors that play a role in successful transfer (size, concentration, and surface charge along with 3 different cell lines: HepG2, HeLa, and Caco-2). We observed that polystyrene (PS) carboxylated NPs with a size of 40 and 100 nm were successfully and efficiently transferred to GPMVs derived from all cell lines. We then investigated the distribution of NPs inside formed GPMVs and established the average number of NPs/GPMVs and the percentage of all GPMVs with NPs in their cavity. We pave the way for GPMV usage as superior cell-like mimics in medically relevant applications.