Regional food systems and organic agriculture are both considered more sustainable than the conventional, globalized food system they provide an alternative to. The emergence and expansion of alternative forms of food supply are influenced by various factors on different scales. Using the food systems approach we aim to study potentials and limitations of regional organic food supply in the Berlin metropolitan region (BMR). Based on the literature, we developed an analytical framework and identified determinants of regional organic food provision along the three major levels of the supply chain: agricultural production, food chain organization, and consumption. Then, we examined a qualitative case study with two different types of alternative food networks (A) organic community supported agriculture (CSA) and (B) organic retail trade. Factors that hinder or promote the provision of regional organic food were identified through qualitative interviews and assessed by regional stakeholders in a workshop. Our findings show that demand for regional organic food is higher than regional supply, which could offer good possibilities for organic farmers. However, actors in these two food chains need to overcome some obstacles, including limited access to land, increasing renting prices, insufficient processing capacities, and unsupportive political environment for organic farming.