Objective Sit-stand workstations have been shown to be effective in reducing sitting time in office workers. The aim of this study was to explore reasons for use and non-use of sit-stand workstations and strategies to decrease sitting and increase physical activity in the workplace from perspectives of users and non-users, as well as from managers and ergo-coaches. Methods Six group interviews with employees who have had access to sit-stand workstations for several years were conducted in a large semi-governmental organisation in the Netherlands. Verbatim transcripts were analysed using thematic analysis. Open coding was conducted by three researchers and codes and themes were discussed within the research team. Results Thematic analysis resulted in two major themes: 1) Reasons for use and non-use and 2) Strategies to increase standing and physical activity in the workplace. Shared and distinct reasons for use and non-use were identified between users and non-users of the sit-stand workstations. The most important reasons for use indicated by users were that they had experiencing immediate benefits, including staying alert and increasing focus; these benefits were not acknowledged by non-users. Non-users indicated that sitting was comfortable for them and that they were therefore not motivated to use the standing option. Strategies to increase the use of the standing option included an introductory phase to become familiar with working while standing and to experience the immediate benefits that come from using the standing option. Furthermore, providing reminders to use the standing option was suggested as a strategy to increase and sustain the use of sit-stand workstations. Increased use may lead to a change in the sitting culture within the organisation, as more employees would adopt active movement behaviours. Conclusion Immediate benefits of the use of the standing option–only mentioned by the users–was the most distinct reason to use sit-stand workstations. Future research should explore how to motivate potential users to adhere to an introductory phase in order to experience these immediate benefits, whether it is linked to the use of sit-stand workstations or other interventions to reduce sitting time.