StemFit For Differentiation (“StemFit Diff”) is a chemically defined, human/animal component-free supplement for optimizing differentiating human embryonic stem (ES) and induced pluripotent stem (iPS) cells. It supports differentiation along ectoderm, mesoderm, or endoderm lineages with various induction factors or cytokines. StemFit Diff can be combined with a basal media (IMDM, DMEM/F12, RPMI1640, etc.) for clinical research applications of human pluripotent stem cell (hPSC)-derived cells/tissues, providing defined and xeno-free culture systems for expansion and differentiation.