Bovine pluripotent stem cell-based muscle organoids, method of preparing the same and uses thereof

A method for generating bovine myogenic progenitor cells, the method comprising:

• (a) culturing bovine pluripotent stem cells (bPSCs) in a first culture medium comprising at least one growth factor for a first time period until aggregates form and the cells express Brachyury, thereby obtaining mesodermal organoids;
• (b) replacing the first culture medium with a second culture medium comprising at least one bone morphogenic protein (BMP) inhibitor and at least one Wnt activator and essentially devoid of a Wnt inhibitor and activin A, and incubating for a second time period until the cells express Tbx6 and Pax3, thereby obtaining anterior paraxial mesodermal organoids and optionally somite-like structures;
• (c) replacing the second culture medium with a third culture medium comprising at least one Wnt activator and at least one hedgehog inhibitor and incubating for a third time period until the cells express MyoD, thereby obtaining myogenic progenitor cells.