We theorize that vicarious learning theory provides a framework for understanding how small- and medium-sized start-ups can learn from the activity of a variety of regional actors, not just from the activity of colocated peer firms (i.e., other start-ups). Furthermore, we suggest that the magnitude of the impact of vicarious learning is influenced by a firm’s own specific experience with a variety of actors. We use longitudinal data of the population of German biotechnology start-ups and pharmaceutical multinational corporations (MNCs) between 1996 and 2015 across 19 German biotechnology regions. We show that colocated start-ups’ international expansion is positively impacted by the regional network centrality of colocated MNCs and that this relationship is moderated by a start-up’s direct alliance experience with these entities. Our results highlight how important it is for researchers to differentiate the distinct and separate influences a wide variety of actors have on vicarious learning to more clearly identify outcomes of this influence. We also provide evidence that the influence of MNCs is heterogeneous and depends on whether MNCs are domestic or foreign and on their R&D intensity, yet find that country of origin has no significant influence. Our study makes a number of contributions, one of which is research on alliances, supporting conflicting arguments on the subsequent impacts of experience. We further find that certain types of alliance experience may not be transferrable to induce start-ups’ future international expansion, and in some cases may even hinder it.