Demand for local food, including strawberries (Fragaria ×ananassa), is increasing throughout the United States. Strawberry production in the midwestern United States can be challenging due to the relatively short growing season and pests. However, vertical, hydroponic, high tunnel production systems could extend the growing season, minimize pest incidence, and maximize strawberry yield and profitability. The objectives of this study were to 1) identify the best cultivars and growing media for vertical, hydroponic, high tunnel production of strawberries in the midwestern United States and to 2) assess potential strategies for replacing synthetic fertilizer with organic nutrient sources in hydroponic strawberry production. To accomplish these objectives, three experiments were conducted across 2 years and two locations in Illinois to compare 11 strawberry cultivars, three soilless media mixtures, and three nutrient sources. Strawberry yield was greatest when grown in perlite mixed with coco coir or vermiculite and fertilized with a synthetic nutrient source. Yield was reduced by up to 15% when fertilized with a bio-based, liquid nutrient source and vermicompost mixed with soilless media. Strawberry yield among cultivars varied by year and location, but Florida Radiance, Monterey, Evie 2, Portola, and Seascape were among the highest-yielding cultivars in at least one site-year. Results contribute to the development of best management practices for vertical, hydroponic, high tunnel strawberry production in the midwestern United States, but further research is needed to understand nutrient dynamics and crop physiological response among levels within vertical, hydroponic towers.