Ebola virus disease outbreaks have repeatedly occurred on the African continent over the last decades, with more serious outbreaks in recent years. Being highly transmissible and associated to high fatality rates, it constitutes a serious threat to public health. Vaccination, however, may allow for efficient control of its propagation. The most promising Ebola vaccine candidate to date, rVSV-ZEBOV, relies on a recombinant vesicular stomatitis virus construct, in which the native viral glycoprotein is replaced by the glycoprotein of Ebola virus (Zaire). However, its cell-based manufacturing process is still lengthy and cumbersome, thus urging the implementation of a new and more efficient bioprocess. To address these issues, serum-free production of rVSV-ZEBOV in Vero cells has been studied with the aim to test an alternative upstream process. Until viable options of suspension cell culture are available, Vero cell cultures still rely on adherent bioprocesses and have thus been developed in this work. Particularly, a bioprocess developed with standard microcarrier bioreactor technology was successfully transferred to the novel single-use scale-X™ hydro fixed-bed.