Preparation of human foreskin fibroblasts for human embryonic stem cell culture

INTRODUCTIONIn order to reduce the risk of immune rejection and zoonosis in human embryonic stem cell (hESC) transplantation recipients, animal-derived components, nonhuman sera, and animal feeder cells need to be removed from hESC culture. To limit the use of animal products, several different human feeder cell types have been tested for hESC culture. These include cells from fetal muscle and skin, as well as adult fallopian tube epithelial cells and adult muscle cells. Human foreskin fibroblasts (hFFs) have been used successfully as feeder cells to support the derivation and undifferentiated growth of hESCs and are commercially available through the American Type Culture Collection (ATCC). This protocol describes a method for preparing hFFs for hESC culture.