The pancreatic ductal adenocarcinoma (PDAC) is the most frequent neoplasm of the pancreas. In vivo pancreatic ductal adenocarcinoma cells are influenced by a number of intra- and extracellular factors through different signalling pathways. Especially pancreatic stellate cells (PSCs) are known to stimulate tumor growth and motility. Here we investigated the influence of PSCs compared to epidermal growth factor (EGF) on PDAC migration, which is an indispensable step for metastasis. We used time-lapse videomicroscopy to capture single cell migration of the PDAC-line BxPC3, while supernatant of the PSC-line RLT-PSC and EGF were added to the media. BxPC3 and RLT-PSC cells were investigated together at the same time in wound healing assays. Cell migration was quantified as the movement of the cell center or as the increase of cell-covered wound area with time, respectively. Migration velocity of PDAC cells was increased by the PSC supernatant and EGF, while the inhibition of the Na⁺/H⁺-exchanger NHE1 reduced the simulating effect of EGF. In wound healing assays the presence of PSCs had no effect on the velocity of BxPC3 cells and BxPC3 cells were not able to increase motility of RLT-PSC cells.The latter observation differs from experiments made with other PDAC cell lines whose supernatant stimulates PSC migration. This points to a PDAC cell-specific composition of the supernatant. In conclusion, migration velocity of single BxPC3 cells is increased by EGF and secreted factors of RLT-PSC cells, while during wound healing movement a stimulation between these cells cannot be observed.