title: Formation and hematopoietic differentiation of human embryoid bodies by suspension and hanging drop cultures.
abstract: The in vitro aggregation of human embryonic stem cells (hESCs) into clusters termed embryoid bodies (EBs) allows for the spontaneous differentiation of cells representing endoderm, mesoderm, and ectoderm lineages. This stochastic process results however, in the generation of low numbers of differentiated cells, and can be enhanced to some extent by the addition of exogenous growth factors or overexpression of regulatory genes. In the authors' laboratory, the use of hematopoietic cytokines in combination with the mesoderm inducer bone morphogenetic protein-4 (BMP-4) was able to generate up to 90% of CD45(+) hematopoietic cells with colony-forming unit (CFU) activity. This unit describes two protocols that have been successfully applied in the authors' laboratory for the generation of EBs in (1) suspension and (2) hanging drop (HD) cultures from enzymatically digested clumps of undifferentiated hESC colonies.
authors.affiliation: McMaster University, Hamilton, Ontario, Canada.
authors.forename: Chantal
authors.initials: C
authors.lastname: Cerdan
authors.forename: Seok Ho
authors.initials: SH
authors.lastname: Hong
authors.forename: Mickie
authors.initials: M
authors.lastname: Bhatia
mesh_terms: D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D016207:Cytokines / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006410:Hematopoiesis / Q000187:drug effects; D006801:Humans; D011994:Recombinant Proteins / Q000494:pharmacology
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D016207:Cytokines; D011994:Recombinant Proteins