This chapter shares examples of organization-level barriers to full participation of women in entrepreneurial ecosystems by way of the three cities that were the sites of our fieldwork—Boston, Massachusetts, St. Louis, Missouri, and Asheville, North Carolina. Here, the focus is on the ways in which intermediary organizations, such as incubators, accelerators, coworking spaces, and investors among others, can act as gatekeepers to the resources of the ecosystem. The chapter focuses specifically on access to networks, outreach, selection, support mechanisms (i.e., entrepreneur support organizations) available in the ecosystem, and ecosystem culture. In speaking to these issues, the chapter focuses on the role of meso-level organizational actors and how their norms, values, and practices differentially impact entrepreneurs and lead to inclusion or exclusion from the ecosystem.