ABSTRACT
This chapter addresses the development of good manufacturing practice compliant production processes for generation of human embryonic stem cell (hESC)-derived differentiated cells. It discusses the manufacturing and regulatory requirements including hESC line qualification, raw materials qualification, process development, cell production, cryopreservation and formulation and product release. Undifferentiated hES cells have been cultured and maintained on mouse embryonic feeder cells in the presence of medium and exogenously added basic fibroblast growth factor. Although the use of a mouse feeder layer allows for long-term maintenance of hES cultures, this culture system results in cells contaminated with mouse feeder cells and is not preferred for research, development or clinical use. The conditions for formulation and cryopreservation are critical for maintaining the function, viability and shelf-life of cells. The parameters that should be considered include freezing medium, cryoprotectants, controlled rate freezing conditions and thawing conditions.
