Abstract
The first haematopoietic stem cells (HSCs) appear in the aorta-gonad-mesonephros (AGM) region, major vitelline and umbilical vessels, and placenta; however, whether they arise locally or from immigrant yolk sac precursor cells remains unclear. This issue is best addressed by measuring cell-lineage relationships rather than cell potentials. To undertake long-term in vivo tracing of yolk sac cells, we designed a non-invasive pulse-labelling system based on Cre/loxP recombination. Here we show that in Runx1+/- (runt-related transcription factor 1) heterozygous mice, yolk sac cells expressing Runx1 at embryonic day 7.5 develop into fetal lymphoid progenitors and adult HSCs. During mid-gestation the labelled (embryonic day 7.5) yolk sac cells colonize the umbilical cord, the AGM region and subsequently the embryonic liver. This raises the possibility that some HSCs associated with major embryonic vasculature are derived from yolk sac precursors. We observed virtually no contribution of the labelled cells towards the yolk sac vasculature, indicating early segregation of endothelial and haematopoietic lineages.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Medvinsky, A. & Dzierzak, E. Definitive hematopoiesis is autonomously initiated by the AGM region. Cell 86, 897–906 (1996)
Gekas, C., Dieterlen-Lievre, F., Orkin, S. H. & Mikolla, H. K. A. The placenta is a niche for hematopoietic stem cells. Dev. Cell 8, 365–375 (2005)
Cumano, A., Dieterlen-Lievre, F. & Godin, I. Lymphoid potential, probed before circulation in mouse, is restricted to caudal intraembryonic splanchnopleura. Cell 86, 907–916 (1996)
Cumano, A., Ferraz, J. C., Klaine, M., Di Santo, J. P. & Godin, I. Intraembryonic, but not yolk sac hematopoietic precursors, isolated before circulation, provide long-term multilineage reconstitution. Immunity 15, 477–485 (2001)
Yoder, M. C. et al. Characterization of definitive lymphohematopoietic stem cells in the day 9 murine yolk sac. Immunity 7, 335–344 (1997)
Weissman, I., Papaioannou, V. & Gardner, R. Fetal Hematopoietic Origins of the Adult Hematolymphoid System (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1978)
Zhang, Y. et al. Inducible site-directed recombination in mouse embryonic stem cells. Nucleic Acids Res. 24, 543–548 (1996)
Okuda, T., Van Deursen, J., Hiebert, S. W., Grosveld, G. & Downing, J. R. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell 84, 321–330 (1996)
Wang, Q. et al. Disruption of the Cbfa2 gene causes necrosis and hemorrhaging in the central nervous system and blocks definitive hematopoiesis. Proc. Natl Acad. Sci. USA 93, 3444–3449 (1996)
North, T. et al. Cbfa2 is required for the formation of intra-aortic hematopoietic clusters. Development 126, 2563–2575 (1999)
Cai, Z. et al. Haploinsufficiency of AML1 affects the temporal and spatial generation of hematopoietic stem cells in the mouse embryo. Immunity 13, 423–431 (2000)
Samokhvalov, I. M. et al. Multifunctional reversible knockout/reporter system enabling fully functional reconstitution of the AML1/Runx1 locus and rescue of hematopoiesis. Genesis 44, 115–121 (2006)
Palis, J., Robertson, S., Kennedy, M., Wall, C. & Keller, G. Development of erythroid and myeloid progenitors in the yolk sac and embryo proper of the mouse. Development 126, 5073–5083 (1999)
Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)
Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001)
Igarashi, H., Kouro, T., Yokota, T., Comp, P. C. & Kincade, P. W. Age and stage dependency of estrogen receptor expression by lymphocyte precursors. Proc. Natl Acad. Sci. USA 98, 15131–15136 (2001)
Kellendonk, C. et al. Inducible site-specific recombination in the brain. J. Mol. Biol. 285, 175–182 (1999)
Brotherton, T. W., Chui, D. H. K., Gauldie, J. & Patterson, M. Hemoglobin ontogeny during normal mouse fetal development. Proc. Natl Acad. Sci. USA 76, 2853–2857 (1979)
Steiner, R. & Vogel, H. On the kinetics of erythroid cell differentiation in fetal mice: I. Microspectrophotometric determination of the hemoglobin content in erythroid cells during gestation. J. Cell. Physiol. 81, 323–338 (1973)
Lien, E. A., Solheim, E. & Ueland, P. M. Distribution of tamoxifen and its metabolites in rat and human tissues during steady-state treatment. Cancer Res. 51, 4837–4844 (1991)
Kisanga, E. R., Gjerde, J., Schjott, J., Mellgren, G. & Lien, E. A. Tamoxifen administration and metabolism in nude mice and nude rats. J. Steroid Biochem. Mol. Biol. 84, 361–367 (2003)
Downs, K. M. & Davies, T. Staging of gastrulating mouse embryos by morphological landmarks in the dissecting microscope. Development 118, 1255–1266 (1993)
Nishikawa, S.-I. et al. In vitro generation of lymphohematopoietic cells from endothelial cells purified from murine embryos. Immunity 8, 761–769 (1998)
Mao, X., Fujiwara, Y. & Orkin, S. H. Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice. Proc. Natl Acad. Sci. USA 96, 5037–5042 (1999)
North, T. E. et al. Runx1 expression marks long-term repopulating hematopoietic stem cells in the midgestation mouse embryo. Immunity 16, 661–672 (2002)
Minot, C. S. Development of the blood, the vascular system and the spleen. In Manual of Human Embryology (eds Keibel, F. and Mall, F. P.) 498–534 (J. B. Lippincott, Philadelphia, 1912)
Gerlai, R. Gene-targeting studies of mammalian behavior: is it the mutation or the background genotype? Trends Neurosci. 19, 177–181 (1996)
Müller-Sieburg, C. E. & Riblet, R. Genetic control of the frequency of hematopoietic stem cells in mice: mapping of a candidate locus to chromosome 1. J. Exp. Med. 183, 1141–1150 (1996)
Young, H. A. et al. Bone marrow and thymus expression of interferon-γ results in severe B-cell lineage reduction, T-cell lineage alterations, and hematopoietic progenitor deficiencies. Blood 89, 583–595 (1997)
Yu, J.-M. et al. Expression of interferon-γ by stromal cells inhibits murine long-term repopulating hematopoietic stem cell activity. Exp. Hematol. 27, 895–903 (1999)
Acknowledgements
We thank S.-i. Aisawa for providing us with TT2 ES cells; F. Costantini for R26R-eYFP mice; N. Kazuki and J. Ure for their help with generating the knock-in mouse strains; and F. Melchers, A. Cumano and members of RIKEN CDB Kobe for critical discussion. I.M.S. was a recipient of a postdoctoral fellowship for foreign researchers from the Japan Society for the Promotion of Science. This work was supported in part by a grant for the Project for Realization of Regenerative Medicine (to S.-i.N.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.
Supplementary information
Supplementary Figures
This file contains Supplementary Figures 1–5 with Legends and detailed Supplementary Methods. The Supplementary Figures present additional data on the modified alleles used in the study as well as the scheme for Mer-Cre-Mer targeting into Runx1 locus. The Supplementary Figures also show the Runx1 expression in E7.5-E8.25 mouse concepti, the highly Runx1-positive cell clusters in day 8 - day 9 yolk sacs and provide the additional information on the analysis of the unspecific cell labeling. Supplementary Figure 5 shows the whole-mount X-Gal staining performed 12 hours after the 4’OHT administration at the nominal stage E8.0. The Supplementary Methods provide details about the Cre knock-in construct and generation of chimeric animals, induction with 4-hydroxitamoxifen, flow cytometry and cell preparations, whole-mount X-Gal staining and cryosectioning and PCR genotyping including the sequences of all primers used in this work. (PDF 4597 kb)
Rights and permissions
About this article
Cite this article
Samokhvalov, I., Samokhvalova, N. & Nishikawa, Si. Cell tracing shows the contribution of the yolk sac to adult haematopoiesis. Nature 446, 1056–1061 (2007). https://doi.org/10.1038/nature05725
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature05725
This article is cited by
-
Circulating macrophages as the mechanistic link between mosaic loss of Y-chromosome and cardiac disease
Cell & Bioscience (2023)
-
Testicular macrophages are recruited during a narrow fetal time window and promote organ-specific developmental functions
Nature Communications (2023)
-
α-Gal Nanoparticles in CNS Trauma: I. In Vitro Activation of Microglia Towards a Pro-Healing State
Tissue Engineering and Regenerative Medicine (2023)
-
Genetic fate-mapping reveals surface accumulation but not deep organ invasion of pleural and peritoneal cavity macrophages following injury
Nature Communications (2021)
-
The Fetal-to-Adult Hematopoietic Stem Cell Transition and its Role in Childhood Hematopoietic Malignancies
Stem Cell Reviews and Reports (2021)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.