title: Direct Induction of Hemogenic Endothelium and Blood by Overexpression of Transcription Factors in Human Pluripotent Stem Cells.
abstract: During development, hematopoietic cells arise from a specialized subset of endothelial cells, hemogenic endothelium (HE). Modeling HE development in vitro is essential for mechanistic studies of the endothelial-hematopoietic transition and hematopoietic specification. Here, we describe a method for the efficient induction of HE from human pluripotent stem cells (hPSCs) by way of overexpression of different sets of transcription factors. The combination of ETV2 and GATA1 or GATA2 TFs is used to induce HE with pan-myeloid potential, while a combination of GATA2 and TAL1 transcription factors allows for the production of HE with erythroid and megakaryocytic potential. The addition of LMO2 to GATA2 and TAL1 combination substantially accelerates differentiation and increases erythroid and megakaryocytic cells production. This method provides an efficient and rapid means of HE induction from hPSCs and allows for the observation of the endothelial-hematopoietic transition in a culture dish. The protocol includes hPSCs transduction procedures and post-transduction analysis of HE and blood progenitors.
authors.affiliation: Primate Research Center, University of Wisconsin-Madison.
authors.forename: Irina
authors.initials: I
authors.lastname: Elcheva
authors.affiliation: Primate Research Center, University of Wisconsin-Madison.
authors.forename: Vera
authors.initials: V
authors.lastname: Brok-Volchanskaya
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Wisconsin School of Medicine and Public Health; islukvin@wisc.edu.
authors.forename: Igor
authors.initials: I
authors.lastname: Slukvin
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D055018:Hemangioblasts / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D014157:Transcription Factors / Q000096:biosynthesis*
chemical_list: D014157:Transcription Factors