Abstract
To identify early populations of committed progenitors derived from human embryonic stem cells (hESCs), we screened self-renewing, BMP4-treated and retinoic acid–treated cultures with >400 antibodies recognizing cell-surface antigens. Sorting of >30 subpopulations followed by transcriptional analysis of developmental genes identified four distinct candidate progenitor groups. Subsets detected in self-renewing cultures, including CXCR4+ cells, expressed primitive endoderm genes. Expression of Cxcr4 in primitive endoderm was confirmed in visceral endoderm of mouse embryos. BMP4-induced progenitors exhibited gene signatures of mesoderm, trophoblast and vascular endothelium, suggesting correspondence to gastrulation-stage primitive streak, chorion and allantois precursors, respectively. Functional studies in vitro and in vivo confirmed that ROR2+ cells produce mesoderm progeny, APA+ cells generate syncytiotrophoblasts and CD87+ cells give rise to vasculature. The same progenitor classes emerged during the differentiation of human induced pluripotent stem cells (hiPSCs). These markers and progenitors provide tools for purifying human tissue-regenerating progenitors and for studying the commitment of pluripotent stem cells to lineage progenitors.
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Acknowledgements
This work was supported by funds provided by the California Institute of Regenerative Medicine (CIRM) (Comprehensive grants RC1-00354-1, RT2-02060). Y.S. was supported by a CDA award from the Human Frontier Science Organization (HFSPO, CDA0063/2007-C). C.T. and A.S.L. are supported by the Howard Hughes Medical Institute Medical Fellows and the Stanford Medical Scholars Program. Y.R. is supported by a Human Frontier Science Program (HFSP) Long Term Fellowship. M.D., C.T. and R.A. are supported by CIRM (Comprehensive grants RC1-00354-1, RT2-02060). We would like to thank C. Chan for pentachrome stainings.
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M.D., I.L.W. and Y.S. designed the experiments and wrote the manuscript. M.D., Y.S., C.T., R.A., Y.R., J.S., A.S.L. and A.R.M. performed experiments and analyzed data. All authors endorse the full content of this work.
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Drukker, M., Tang, C., Ardehali, R. et al. Isolation of primitive endoderm, mesoderm, vascular endothelial and trophoblast progenitors from human pluripotent stem cells. Nat Biotechnol 30, 531–542 (2012). https://doi.org/10.1038/nbt.2239
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DOI: https://doi.org/10.1038/nbt.2239
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