The hallmark of pluripotent stem cells is their nearly unlimited self-renewal capacity, and their potential to differentiate into a diverse range of specialized cell types. These unique properties make stem cells important research tools, in vitro models for pharmaceutical testing, and an attractive source of various cell types for regenerative therapies. For stem cell technology to be fully exploited, however, culture systems must be improved to enable large-scale production, and safety ensured. Most stem cell culture systems developed to date utilize undefined, xenogeneic products that pose a risk of a severe immune response and the transmission of infections. In this chapter, we describe a robust method for the expansion of pluripotent stem cells in defined and xeno-free culture conditions. Both mechanical and single-cell enzymatic passaging can be applied with this method. This procedure can be adopted for both basic research purposes and clinical applications.
