Low Cost, Chemically Defined, GM-Free Animal iPSC Generation, Culture, and Skeletal Muscle Differentiation

Common iPSC culture and differentiation media typically cost $500 per L, preventing the application of this technology to the cultivated meat field. To produce whole-cut meat, a low-cost source of high-quality non-genetically modified (GM) animal skeletal muscle cells is essential. Here we will (1) optimize chemically defined media protocols for the non-GM generation and low- cost long-term culture of porcine and bovine iPSC. We will also (2) generate advanced, efficient chemically defined, small molecule-based protocols for differentiation of these porcine and bovine iPSC to skeletal muscle. Finally, we will (3) develop skeletal muscle maturation protocols to improve the bulk and organoleptic properties of these cells, adapt the differentiation protocol to 2.7 liter bioreactors for scalability, and assess seeding of protein scaffolds in a perfusion tissue bioreactor to produce whole-cut meat.