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Stem Cells. 2016 May;34(5):1239-50. doi: 10.1002/stem.2351. Epub 2016 Mar 28.

Genetic Tagging During Human Mesoderm Differentiation Reveals Tripotent Lateral Plate Mesodermal Progenitors.

Author information

1
Department of Pathology & Laboratory Medicine, David Geffen School of Medicine (DGSOM).
2
Molecular Biology Interdepartmental PhD Program, DGSOM University of California Los Angeles.
3
Department of Microbiology, Immunology and Molecular Genetics, DGSOM, DGSOM University of California Los Angeles.
4
Department of Orthopedic Surgery, Keck School of Medicine of University of Southern California (USC). All in, Los Angeles, CA, United States.
5
Department of Molecular, Cell and Development Biology, DGSOM University of California Los Angeles.
6
Molecular Biology Institute (MBI).
7
Department of Pediatrics, DGSOM University of California Los Angeles.
8
Broad Stem Cell Research Center (BSCRC), DGSOM University of California Los Angeles.
9
Jonsson Comprehensive Cancer Center (JCCC), DGSOM University of California Los Angeles.
10
Department of Pathology & Laboratory Medicine, DGSOM University of California Los Angeles.

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.

KEYWORDS:

Clonal tracking; Hematopoiesis; Lentiviral vectors; Lineage tracing; Mesoderm; Pluripotent stem cells; Vector integration site analysis

PMID:
26934332
PMCID:
PMC5052131
DOI:
10.1002/stem.2351
[Indexed for MEDLINE]
Free PMC Article

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