A defined, feeder-independent medium for human embryonic stem cell culture

Abstract

The developmental potential of human ES cells makes them an important tool in develop- mental, pharmacological, and clinical research. For human ES cell technology to be fully exploited, however, culture efficiency must be improved, large-scale culture enabled, and safety ensured. Traditional human ES cell culture systems have relied on serum products and mouse feeder layers, which limit the scale, present biological variability, and expose the cells to potential contaminants. Defined, feeder-independent culture systems improve the safety and efficiency of ES cell technology, enabling translational research. The pro- tocols herein are designed with the standard research laboratory in mind. They contain recipes for the formulation of mTeSR (a defined medium for human ES cell culture) and detailed protocols for the culture, transfer, and passage of cells grown in these feeder- independent conditions. They provide a basis for routine feeder-independent culture, and a starting point for additional optimization of culture conditions.