The derivation of human embryonic stem (hES) cells is a challenging procedure. The isolation and maintenance of hES is visually and manually complicated, involving mechanical or enzymatic passaging using either collagenase or trypsin. This chapter describes detailed protocols that have been used for the derivation, maintenance, and characterization of hES cells in vitro along with protocols to test their differentiation potential in vivo. When used as a guideline, these protocols will assist researchers in setting up a hES cell laboratory.
