Yeasts belonging to the Malassezia genus represent the predominant fungi of the human skin microbiota. Their involvement in various pathologies is notably associated with their ability to form biofilms. However, knowledge regarding these structures remains limited due to culture constraints related to lipid dependence. This study aimed to develop a reliable in vitro model for studying mono- and multispecies Malassezia biofilms, using M. furfur, M. pachydermatis, M. sympodialis , and M. slooffiae . To overcome the natural aggregation of cells, a sonication protocol was optimized to obtain homogeneous suspensions without affecting cell viability. A correlation between optical density at 600 nm and cell concentration enabled the rapid standardization of Malassezia inocula. The comparative study of planktonic growth kinetics revealed generation times ranging from 7 to 15 h depending on the species, with M. sympodialis exhibiting the fastest growth. No difference was observed for M. furfur and M. sympodialis grown in single or mixed culture in planktonic conditions. Biofilm development was monitored over 168 h by quantifying biomass and metabolic activity. M. furfur, M. pachydermatis , and M. sympodialis showed a significant increase in both parameters, typical of mature biofilms. In contrast, M. slooffiae exhibited an increase in metabolic activity without a significant increase in biomass, suggesting an unusual biofilm phenotype requiring further investigation. This study proposes a standardized biofilm model for the evaluation of mono- and multispecies biofilms suitable for the different species of Malassezia . This model represents a relevant tool for studying interspecies interactions and for the development of new antifungal strategies targeting Malassezia biofilms.