Protocol to generate endothelial cells, pericytes, and fibroblasts in one differentiation round from human- induced pluripotent stem cells

Here, we present a protocol for differentiating human-induced pluripotent stem cells into three distinct mesodermal cell types: vascular endothelial cells (ECs), pericytes, and fibroblasts. We describe steps for using monolayer serum-free dif-ferentiation and isolating ECs (CD31+) and mesenchymal pre-pericytes (CD31-) from a single differentiation set. We then differentiate pericytes into fibroblasts using a commercial fibroblast culture medium. The three cell types differentiated in this protocol are useful for vasculogenesis, drug testing, and tissue engineer-ing applications. For complete details on the use and execution of this protocol, please refer to Orlova et al. (2014).1