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Cell tracing shows the contribution of the yolk sac to adult haematopoiesis


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.

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Figure 1: Embryonic expression of Runx1 and analysis of inducible Runx1 -dependent cell tagging.
Figure 2: Embryo cell tagging kinetics at E7.5.
Figure 3: The embryonic haematopoietic progeny of yolk sac blood-island cells.
Figure 4: Representative β-gal staining of E10.5 dorsal aorta and E11.5 umbilical artery and vein sections of embryos activated at E7.5 by a single injection of 4′OHT.
Figure 5: The haematopoietic progeny in adult mice of yolk sac cells from the blood-island region.

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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.

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Correspondence to Igor M. Samokhvalov.

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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)

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Samokhvalov, I., Samokhvalova, N. & Nishikawa, Si. Cell tracing shows the contribution of the yolk sac to adult haematopoiesis. Nature 446, 1056–1061 (2007).

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