The world population is predicted to grow to 10 billion by 2050 that will lead to many food production challenges present significant sustainability and security threats. Cellular agriculture is emerging as an advantageous solution to address these immediate problems by developing sustainable alternative food systems to feed a rapidly growing global population with nutritious and safe food, while protecting our environment and limited resources. Our lab is focused on growing seafood products, specifically fish, that require less resources compared to mammalian cell culture, making them good targets for cultivated meat. However, current cellular agriculture expansion is limited by serum-containing media used to grow cells . The serum component is expensive, unreliably produced, and questionably sourced making it important to create cost-effective and chemically defined serum-free. To our knowledge, there is no published serum-free media formulation specifically designed for long-term growth of food-grade fish cell lines.
Our lab is developing a multi-leveled machine learning protocol to optimize chemically defined serum-free media development for lean fish cell lines with verified growth factors in the least number of possible experimentations. Our first model designed 93 experiments to grow Zebrafish embryonic stem cells with seven component variables at five concentration levels. This model found that traditional growth factors and components used in mammalian serum-free cell culture are not all suitable for fish cells, and that serum remains the most stable component in growth. However, serum was not the most critical component in the initial days of growth compared to more traditionally used growth factors (Fig. 1) . Current work optimizing the model with alternative growth factors that may be more crucial for fish cells , such as the essential fatty acids, to continue decreasing serum levels in growth media. Additional cultivated meat related responses will be measured for the best growth medias including cryopreservation viability , proliferation potential, differentiation potential, and chromosome stability. Lastly, the most propitious growth medias will be scaled-up in our mini-bioreactor to further validate the growth media formulation. This methodology will create a durable predictive model tool applicable to developing serum-free media for alternative fish cellular agriculture cell lines and increase the capacity of cultivated food on an industrial scale.