Encouraging the use of collaboration in regional economic development has been increasingly prevalent over the past few decades by both governmental and non-governmental stakeholders. This push has two origins. First, regional level stakeholders are coming to understand the limits of what they can achieve as individual organizations. This has made collaboration at a regional level attractive. Second, provincial, state and federal levels of government increasingly prefer to interact with only one entity at a regional level, making consolidation of effort through collaboration key. There is an expectation that regional collaborations between multiple cities and counties will help to mobilize more resources, than a single municipality would be able to do alone. This study explores collaborations that connect diverse stakeholders (public officials, private organizations, and non-governmental entities) related to regional agriculture initiatives and regional economic development in the North Country in New York state and Four Counties in Ontario. Both regions have strong agricultural economies, are vast areas geographically, and have low population densities. Thirty-four semi-structured interviews were conducted, focusing on factors key in success or failure; barriers to cooperation between different organizations and communities; and challenges in implementing regional development initiatives. The capacity of the case regions to participate in regional collaboration has been varied. To date, regional collaboration has been more developed in the North Country compared to the Four Counties, primarily due to the state organized regional council process that has given the region structure for its economic development initiatives, as well as a deeply embedded history of acting as a region. Key factors for success in economic development collaboration in these two case study areas included; utilizing communication technology, embracing regional attributes, diverse membership, flexible structure, appropriate use of leadership, and insufficient financial capital. Barriers to collaborating included; communication technology, diverse membership, outlook toward collaboration, insufficient human capital, and insufficient financial capital. Therefore, several factors were seen to be necessary for success, but also barriers depending on the regional context. With the exception of the contradictory role of technology, these findings are supported by the literature. This study also identifies several benefits highlighted by county level stakeholders of not collaborating at a regional scale, such as enhanced relationship building with local producers and lower-tier municipalities, and avoiding unnecessary layers of bureaucracy. This study reveals that regional collaboration is not a universal solution to economic challenges for all jurisdictions. Collaboration at an intra county scale may be more desirable and successful. This study also identifies several characteristics that facilitate collaboration that are not in line with the literature including a vast geography, no urban centre, and insufficient resources. This is significant, as these characteristics are often considered barriers to economic development and collaboration. This study serves to expand the literature on experiences of collaboration in rural regions; enhancing the understanding of disadvantaged regions' experiences with economic development; and broadening the analysis of collaboration to not only include government actors, but also not-for-profit and for-profit businesses, non- governmental organizations, and civil society organizations.