StemFit® For Differentiation (“StemFit® Diff”) is a chemically defined and human/animal component-free supplement for differentiation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells to multiple lineages. It can be used with a variety of different induction factors or cytokines to support differentiation along ectoderm, mesoderm or endoderm lineages. StemFit® Diff can be combined with StemFit® Basic media, using iMatrix-511 laminin as extracellular matrix, to support clinical research applications of human pluripotent stem cell (hPSC)-derived cells/tissues by providing defined and xeno-free culture systems for both expansion and differentiation. StemFit® Diff is provided as 5X concentrate and is intended to be used with basal medium (DMEM, DMEM/F12, RPMI1640, etc. and appropriate induction factors or cytokines.