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Germ-layer and lineage-restricted stem/progenitors regenerate the mouse digit tip

Nature volume 476, pages 409–413 (2011)Cite this article

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Abstract

The regrowth of amputated limbs and the distal tips of digits represent models of tissue regeneration in amphibians, fish and mice. For decades it had been assumed that limb regeneration derived from the blastema, an undifferentiated pluripotent cell population thought to be derived from mature cells via dedifferentiation. Here we show that a wide range of tissue stem/progenitor cells contribute towards the restoration of the mouse distal digit. Genetic fate mapping and clonal analysis of individual cells revealed that these stem cells are lineage restricted, mimicking digit growth during development. Transplantation of cyan-fluorescent-protein-expressing haematopoietic stem cells, and parabiosis between genetically marked mice, confirmed that the stem/progenitor cells are tissue resident, including the cells involved in angiogenesis. These results, combined with those from appendage regeneration in other vertebrate subphyla, collectively demonstrate that tissue stem cells rather than pluripotent blastema cells are an evolutionarily conserved cellular mode for limb regeneration after amputation.

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Figure 1: Germ-layer restriction of ectoderm/mesoderm during digit-tip regeneration.
Figure 2: Lineage restrictions of bone, tendons and endothelium during digit-tip regeneration.
Figure 3: Circulating cells do not contribute to regenerating tissues of the digit tip.
Figure 4: Multiple lineage-restricted clones contribute to digit-tip regeneration.

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Acknowledgements

We thank A. Mosley for assistance with animal care and parabiosis experiments; A. Sweet-Cordero for providing Sox9Cre and Prx1Cre mice; R. Schweitzer for providing ScxCre mice; and L. Luo for providing mTmG reporter mice. We thank D. Montoro for assistance with histochemical stains and G. Paz for assistance with figure preparations. This work was supported in part by a grant from the California Institute of Regenerative Medicine (RC1 00354) and from the Smith Family Trust (to I.L.W.), the Oak Foundation and the Hagey Laboratory for Pediatric Regenerative Medicine (to M.T.L.). Y.R. is supported by a Human Frontier Science Program (HFSP) Long Term Fellowship, and the Machiah Foundation Fellowship.

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Author notes

  1. Yuval Rinkevich

    Present address: Present address: Institute for Stem Cell Biology and Regenerative Medicine, Lorry I. Lokey Research Building, Stanford University, Stanford, California 94028, USA.,

Authors and Affiliations

  1. Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, 94305-5323, California, USA

    Yuval Rinkevich, Paul Lindau, Hiroo Ueno, Michael T. Longaker & Irving L. Weissman

  2. Department of Surgery, Hagey Laboratory for Pediatric Regenerative Medicine, Plastic and Reconstructive Surgery, Stanford University School of Medicine, Stanford, 94305, California, USA

    Michael T. Longaker

  3. Ludwig Center for Cancer Stem Cell Research, Stanford University, Stanford, 94305-5323, California, USA

    Irving L. Weissman

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Contributions

Y.R. and I.L.W. designed the experiments. Y.R. performed the regeneration experiments, imaged and analysed the data from all regeneration experiments. H.U. provided the Rainbow reporter mice for part of the experiments. M.T.L. provided the histochemical stains for part of the experiments. Y.R. and P.L. performed the HSC transplantations and analysed the data. Y.R. and I.L.W. wrote the manuscript.

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Correspondence to Yuval Rinkevich.

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The authors declare no competing financial interests.

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Rinkevich, Y., Lindau, P., Ueno, H. et al. Germ-layer and lineage-restricted stem/progenitors regenerate the mouse digit tip. Nature 476, 409–413 (2011). https://doi.org/10.1038/nature10346

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