Conventionally, human embryonic stem cells (hESCs) are cultured on feeder cells. The most commonly used feeder cells are mouse embryonic fibroblasts. It is thought that the feeder cells provide an optimal microenvironment for the undifferentiated growth of hESCs by supplying currently unidentified extracellular matrix components and growth factors. Although these feeder cells are efficient, the feeder-dependent hESC culture system may not be suitable for drug screening or other research purposes that require the use of feeder-free culture methods. In this chapter, we describe in detail protocols for three widely used feeder-free systems for culturing hESCs: Matrigel-mTeSR1, CELLstart-STEMPRO� hESC SFM, and vitronectin-mTeSR1.
