Center for iPS cell research and application (CiRA) 1 article published in JoVE Developmental Biology Hemogenic Endothelium Differentiation from Human Pluripotent Stem Cells in A Feeder- and Xeno-free Defined Condition Ryo Ohta*1, Ryohichi Sugimura*1, Akira Niwa1, Megumu K. Saito1 1Center for iPS cell research and application (CiRA) Here, we present a protocol to precisely form hemogenic endothelium from human pluripotent stem cells in a feeder- and xeno-free condition. This method will have broad applications in disease modeling and screening for therapeutic compounds.