The present study focuses on the design, production and characterization of lipid-based nanosystems as a tool to deliver manganese for diagnostic purpose in multimodal imaging techniques. Particularly, aqueous dispersions of anionic liposomes have been considered as delivery systems for different manganese-based compounds. Negatively charged liposomes were obtained by direct hydration method followed by extrusion using in their composition different anionic surfactants, such as N-lauroylsarcosine (NLS) and sodium lauroyl lactylate (SLL). After production, liposomes were characterized in terms of size, polydispersity, surface charge and stability over time. The extruded liposome dispersions were homogeneous, monodispersed (with an average diameter not exceeding 200 nm) and negatively charged as confirmed by ζ potential measurement. Moreover, as indicated by atomic absorption spectroscopy analyses, the loading of manganese-based compounds was almost quantitative. Liposomes composed of NLS or SLL were stable over time and the presence of manganese-based compounds did not affect their size distribution. In addition their magnetic properties have been retained. The in vitro cytotoxicity of the prepared anionic liposomes was evaluated by MTT assay on human keratinocyte. The obtained results highlighted that unloaded formulations and liposomes loaded with hydrophilic manganese derivatives do not affect cell viability, while liposomes loaded with lipophilic manganese derivatives showed a dose-dependent antiproliferative effect. Therefore further experiments need to be performed in order to elucidate the optimal type and concentration of manganese derivative evaluating both the loading capacity and its bioavailability possibly using other non-tumor human cell lines, such as fibroblasts.