title: Clump passaging and expansion of human embryonic and induced pluripotent stem cells on mouse embryonic fibroblast feeder cells.
abstract: The ability of human embryonic stem cells (hESCs) to differentiate into essentially all somatic cell types has made them a valuable tool for studying human development and has positioned them for broad applications in toxicology, regenerative medicine, and drug discovery. This unit describes a protocol for the large-scale expansion and maintenance of hESCs in vitro. hESC cultures must maintain a balance between the cellular states of pluripotency and differentiation; thus, researchers must use care when growing these technically demanding cells. The culture system is based largely on the use of a proprietary serum-replacement product and basic fibroblast growth factor (bFGF), with mouse embryonic fibroblasts as a feeder layer. These conditions provide the basis for relatively inexpensive maintenance and expansion of hESCs, as well as their engineered counterparts, human induced pluripotent stem cells (hiPSCs).
authors.affiliation: Stem Cell Program, Children's Hospital Boston, Boston, Massachusetts, USA.
authors.forename: Odelya
authors.initials: O
authors.lastname: Hartung
authors.forename: Hongguang
authors.initials: H
authors.lastname: Huo
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
authors.forename: Thorsten M
authors.initials: TM
authors.lastname: Schlaeger
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods; D004622:Embryo, Mammalian; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005333:Fetus / Q000166:cytology* / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology; D051379:Mice; D008954:Models, Biological; D018874:Spheroids, Cellular / Q000166:cytology* / Q000502:physiology