The MesoTotal™ HPC/HSC differentiation system is a new medium and protocol for high efficiency differentiation of human pluripotent stem cells into hematopoietic progenitor cells (CD45/43+ and CD34+) and hematopoietic stem cells (CD45/43+, CD34+, CD38, CD45RA, CD90+). Yields can exceed the publication record by 100-fold and 40-fold for HPCs and HSCs, respectively. The MesoTotal differentiation system involves a two-step incubation procedure, eliminates feeder cell co-culturing, and can improve yields from good blood-producing cell lines.