Human pluripotent stem cells (hPSCs) have the ability to generate all cell types in the human body and can be used in many applications in basic research and translational medicine, including disease modeling, drug screening and cell therapy. Maintenance of high quality hPSCs is dependent on consistent in vitro cell culture conditions and handling techniques. TeSR™-E8™ is a defined, xeno-free medium with a simple and published formula that was developed to meet the demands of a wide range of applications. The E8 culture system has been successfully used to derive and maintain hPSCs, and enzyme-free methods have also been developed for cell expansion and cryopreservation. More recently, defined differentiation conditions have also been established for specific lineages. These developments will provide an efficient and cost-effective E8 platform for the hPSC field.