A Quantitative Proteomic Analysis of Hemogenic Endothelium Reveals Differential Regulation of Hematopoiesis by SOX17.
The in vitro derivation of hematopoietic stem cells (HSCs) from pluripotent stem cells (PSCs) is complicated by the existence of multiple overlapping embryonic blood cell programs called primitive, erythromyeloid progenitor (EMP), and definitive. As HSCs are only generated during the definitive stage of hematopoiesis, deciphering the regulatory pathways that control the emergence of this program and identifying markers that distinguish it from the other programs are essential. To identify definitive specific pathways and marker sets, we used label-free proteomics to determine the proteome of embryo-derived and mouse embryonic stem cell-derived VE-CADHERIN(+)CD45(-) definitive hematopoietic progenitors. With this approach, we identified Stat1 as a marker that distinguishes the definitive erythroid lineage from the primitive- and EMP-derived lineages. Additionally, we provide evidence that the generation of the Stat1(+) definitive lineage is dependent on Sox17. These findings establish an approach for monitoring the emergence of definitive hematopoiesis in the PSC differentiation cultures.
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, ON M5G 1L7, Canada.
Raedun L
RL
Clarke
Institute for Stem Cell and Regenerative Medicine and Department of Pharmacology, University of Washington School of Medicine, Seattle, WA 98109, USA.
Aaron M
AM
Robitaille
Institute for Stem Cell and Regenerative Medicine and Department of Pharmacology, University of Washington School of Medicine, Seattle, WA 98109, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA. Electronic address: rtmoon@uw.edu.
Randall T
RT
Moon
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, ON M5G 1L7, Canada. Electronic address: gkeller@uhnresearch.ca.
Gordon
G
Keller
D000818:Animals; D019070:Cell Lineage; D002478:Cells, Cultured; D066026:Endothelial Progenitor Cells / Q000166:cytology* / Q000378:metabolism; D024002:HMGB Proteins / Q000235:genetics / Q000378:metabolism*; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D020543:Proteome; D055760:SOXF Transcription Factors / Q000235:genetics / Q000378:metabolism*; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism
D024002:HMGB Proteins; D020543:Proteome; D055760:SOXF Transcription Factors; D050794:STAT1 Transcription Factor; C511606:Sox17 protein, mouse; C494080:Stat1 protein, mouse