Through their pollination services, honey bees serve an essential role in sustaining the nutrition, health, and shelter of humans and other animals. Because of the demand for their pollination services, beekeepers strive to keep honey bees healthy and productive. Unfortunately, many factors commonly found throughout the USA can detract from honey bee health. For example, infections with pathogens such as viruses and Nosema spp. pose a significant threat to honey bee survival. Viruses and Nosema spp. must invade cells of their host to reproduce and complete their development. To better understand the interaction between intracellular pathogens and honey bee cells, tools are needed that allow examination of infection at fine resolution and under controlled, aseptic conditions. One tool that has been underutilized in honey bee biology is cell culture; therefore, the focus of this chapter is to discuss the technique and use of honey bee cells in culture. Infection of honey bee cells in culture with the fungal pathogen, Nosema ceranae, is used as an example to demonstrate how cell culture can be a powerful tool to explore the process of infection and the negative impact pathogens may have on honey bee biology and health.