Self-organization of vascularized muscle from bovine embryonic stem cells

Cultured beef offers a promising alternative to traditional meat. While adult stem cells are commonly used as the cell source for cultured beef, their proliferation and differentiation capacities are limited. Current manufacturing processes for cultured beef steaks often require separate preparation of multiple cell types and their intricate assembly. In this study, we propose and report the co-induction of muscle, neural, and endothelial cells from bovine embryonic stem cells (ESCs), accompanied by the formation of tissue-like structures in two- and three-dimensional cultures. Bovine myocytes are induced in a stepwise manner through the induction of presomitic mesoderm (PSM) from bovine ESCs. Muscle fibers with sarcomeres appear within 15 days, displaying calcium oscillations responsive to inputs from co-induced bovine neurons. Bovine endothelial cells are also co-induced via PSM, forming uniform vessel networks inside tissues. Our serum-free, rapid co-induction protocols represent a milestone toward self-organizing beef steaks with integrated vasculature and innervation.