Multilineage cardiovascular organoids and methods of generating the same

The multilineage cardiovascular organoid of any one of the preceding claims, wherein the non-mesodermal cell types comprise endoderm epithelial cells, liver cells, intestinal cells, and/or cardiac neural crest cells.

The multilineage cardiovascular organoid of any one of the preceding claims, wherein the mesodermal cell types comprise cardiac progenitor cells, proliferating cells, cardiomyocytes, and fibroblasts, and wherein the non-mesodermal cell types comprise endoderm epithelial cells, liver cells, intestinal cells, and cardiac neural crest cells.

The multilineage cardiovascular organoid of any one of the preceding claims, wherein the mesodermal and non-mesodermal cell types are determined by expression of one or more markers for each cell type.

The multilineage cardiovascular organoid of any one of the preceding claims, for use in a method of assessing cardiotoxicity of an agent or condition.

A method of generating a multilineage cardiovascular organoid, the method comprising:

• a. culturing an embryoid body in a cardiac induction medium comprising a Wnt signaling activator;
• and
• b. culturing the embryoid body in a Wnt inhibitor medium comprising a Wnt signaling inhibitor.

A method of generating a multilineage cardiovascular organoid, comprising:

• a. generating an embryoid body from induced pluripotent stem cells (iPSCs);
• and
• b. guiding differentiation of the embryoid body to a cardiac lineage, wherein guiding differentiation of the embryoid body to a cardiac lineage comprises culturing an embryoid body in a cardiac induction medium comprising a Wnt signaling activator.

A system comprising a plurality of multilineage cardiovascular organoids, wherein each of the plurality of multilineage cardiovascular organoids comprises mesodermal and non-mesodermal cell types.