As concepts for incorporating uncrewed aerial vehicles (UAS) into controlled airspace are being developed, the need for automated UAS traffic management (UTM) systems to guide UAS and maintain safety is becoming more apparent. A major point of concern for the implementation of UTM is how such systems could coexist alongside the human-centric air traffic management system that is already in place. The European Union’s U-space concept proposes the use of dynamic segregation of airspace reserved for UAS within the control zone. We conducted a simulation experiment with ten air traffic control officer (ATCO) volunteers to gather insights into the feasibility of tower controllers performing the dynamic segregation task. An interface prototype that supports dynamic geofencing and low-level UAS control was developed for this purpose. We found that our proposed interface design helped ATCOs detect potential conflicts between UAS and crewed aircraft. However, they were not always able to adequately resolve them, which resulted in several loss of separation events. It appears that the limitations of the dynamic segregation concept do not fit well with typical air traffic control strategies used by ATCOs. To substantiate our findings, we propose future research to investigate how to overcome the limitations of dynamic segregation to resolve tactical conflicts by revising ATCO control strategies, reevaluating their role in dynamic segregation, as well as considering the definition of flight rules and separation minima for UAS.