title: Genetic Tagging During Human Mesoderm Differentiation Reveals Tripotent Lateral Plate Mesodermal Progenitors.
abstract: Although clonal studies of lineage potential have been extensively applied to organ specific stem and progenitor cells, much less is known about the clonal origins of lineages formed from the germ layers in early embryogenesis. We applied lentiviral tagging followed by vector integration site analysis (VISA) with high-throughput sequencing to investigate the ontogeny of the hematopoietic, endothelial and mesenchymal lineages as they emerge from human embryonic mesoderm. In contrast to studies that have used VISA to track differentiation of self-renewing stem cell clones that amplify significantly over time, we focused on a population of progenitor clones with limited self-renewal capability. Our analyses uncovered the critical influence of sampling on the interpretation of lentiviral tag sharing, particularly among complex populations with minimal clonal duplication. By applying a quantitative framework to estimate the degree of undersampling we revealed the existence of tripotent mesodermal progenitors derived from pluripotent stem cells, and the subsequent bifurcation of their differentiation into bipotent endothelial/hematopoietic or endothelial/mesenchymal progenitors. Stem Cells 2016;34:1239-1250.
authors.affiliation: Department of Pathology & Laboratory Medicine, David Geffen School of Medicine (DGSOM).
authors.forename: Chee Jia
authors.initials: CJ
authors.lastname: Chin
authors.affiliation: Molecular Biology Interdepartmental PhD Program, DGSOM University of California Los Angeles.|Department of Microbiology, Immunology and Molecular Genetics, DGSOM, DGSOM University of California Los Angeles.
authors.forename: Aaron R
authors.initials: AR
authors.lastname: Cooper
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, DGSOM, DGSOM University of California Los Angeles.
authors.forename: Georgia R
authors.initials: GR
authors.lastname: Lill
authors.affiliation: Department of Orthopedic Surgery, Keck School of Medicine of University of Southern California (USC). All in, Los Angeles, CA, United States.
authors.forename: Denis
authors.initials: D
authors.lastname: Evseenko
authors.affiliation: Department of Pathology & Laboratory Medicine, David Geffen School of Medicine (DGSOM).
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Pathology & Laboratory Medicine, David Geffen School of Medicine (DGSOM).
authors.forename: Chong Bin
authors.initials: CB
authors.lastname: He
authors.affiliation: Department of Pathology & Laboratory Medicine, David Geffen School of Medicine (DGSOM).
authors.forename: David
authors.initials: D
authors.lastname: Casero
authors.affiliation: Department of Molecular, Cell and Development Biology, DGSOM University of California Los Angeles.|Molecular Biology Institute (MBI).
authors.forename: Matteo
authors.initials: M
authors.lastname: Pellegrini
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, DGSOM, DGSOM University of California Los Angeles.|Molecular Biology Institute (MBI).|Department of Pediatrics, DGSOM University of California Los Angeles.|Broad Stem Cell Research Center (BSCRC), DGSOM University of California Los Angeles.|Jonsson Comprehensive Cancer Center (JCCC), DGSOM University of California Los Angeles.
authors.forename: Donald B
authors.initials: DB
authors.lastname: Kohn
authors.affiliation: Department of Pathology & Laboratory Medicine, David Geffen School of Medicine (DGSOM).|Department of Pediatrics, DGSOM University of California Los Angeles.|Broad Stem Cell Research Center (BSCRC), DGSOM University of California Los Angeles.|Department of Pathology & Laboratory Medicine, DGSOM University of California Los Angeles.
authors.forename: Gay M
authors.initials: GM
authors.lastname: Crooks
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002469:Cell Separation; D002999:Clone Cells; D005434:Flow Cytometry; D005821:Genetic Techniques; D006801:Humans; D016086:Lentivirus / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology*; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology*
chemical_list: D015703:Antigens, CD
keywords: Clonal tracking; Hematopoiesis; Lentiviral vectors; Lineage tracing; Mesoderm; Pluripotent stem cells; Vector integration site analysis