Honey bee (Apis mellifera) research has been limited by a scarcity of resources for molecular biology. Cell lines in particular are important tools for studying, e.g., signalling cascades or host-pathogen interactions; however, only one stable honey bee cell line has been reported in the literature¹ and it is not widely used. Here we provide proteomic characterization of honey bee primary cells which can be maintained in cell culture for up to one year. The cells are a mixed population derived from worker pupae, so we were first interested in determining what proteins are regularly expressed and what organ or tissue type has the most overlap in expression profile. In this analysis, we identified 2,498 protein groups and the expression profile unequivocally overlaps with the thoracic salivary gland. We are now developing a lipid nanoparticle transfection method and are exploring the utility of the T2A self-cleaving peptide system in these cells. The T2A sequence can be used to liberate protein concatamers, allowing for expression of multiple proteins from one promoter. This system has been shown to be effective in Sf21 cell extracts for in vitro translation,² but has never before been investigated in the honey bee. The work described here represents important developments in the biochemical tools available to honey bee researchers, and will facilitate further research on this important insect.