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Generation of a functional mammary gland from a single stem cell

Abstract

The existence of mammary stem cells (MaSCs) has been postulated from evidence that the mammary gland can be regenerated by transplantation of epithelial fragments in mice1,2,3. Interest in MaSCs has been further stimulated by their potential role in breast tumorigenesis4. However, the identity and purification of MaSCs has proved elusive owing to the lack of defined markers. We isolated discrete populations of mouse mammary cells on the basis of cell-surface markers and identified a subpopulation (Lin-CD29hiCD24+) that is highly enriched for MaSCs by transplantation. Here we show that a single cell, marked with a LacZ transgene, can reconstitute a complete mammary gland in vivo. The transplanted cell contributed to both the luminal and myoepithelial lineages and generated functional lobuloalveolar units during pregnancy. The self-renewing capacity of these cells was demonstrated by serial transplantation of clonal outgrowths. In support of a potential role for MaSCs in breast cancer, the stem-cell-enriched subpopulation was expanded in premalignant mammary tissue from MMTV-wnt-1 mice and contained a higher number of MaSCs. Our data establish that single cells within the Lin-CD29hiCD24+ population are multipotent and self-renewing, properties that define them as MaSCs.

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Figure 1: Enrichment of mammary repopulating units in the Lin - CD29 hi CD24 + population.
Figure 2: Characterization of cytokeratin expression in Lin- cells and increased progenitor capacity of Lin-CD29hiCD24+ mammary cells in vitro.
Figure 3: A single, self-renewing Lin - CD29 hi CD24 + cell can repopulate a MFP.
Figure 4: The Lin - CD29 hi CD24 + population is expanded in MMTV- wnt-1 transgenic mice.

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Acknowledgements

We are grateful to N. Forrest for expert assistance, J. Adams and A. Harris for critical review of the manuscript, S. Mihajlovic for histology, and F. Battye and A. Holloway for FACS support. This work was supported by the Victorian Breast Cancer Research Consortium and the National Health and Medical Research Council (Australia). M.-L.A.-L. is supported by a Fondation pour la Recherche Medicale Fellowship, K.J.S. by a Peter Doherty Fellowship and J.S. by Fellowships from the Canadian Breast Cancer Foundation and the Natural Sciences and Engineering Research Council of Canada. Author Contributions M.S. and F.V. contributed equally to this work. G.J.L. and J.E.V. contributed equally to this work.

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Correspondence to Geoffrey J. Lindeman or Jane E. Visvader.

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Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Methods

This file contains additional details of the methods used in this study. (DOC 27 kb)

Supplementary Figure Legends

This file contains the figure legends for the three Supplementary Figures (DOC 31 kb)

Supplementary Figure 1

Enrichment of long-term label-retaining cells (LT-LRCs) in the Lin-CD29hiCD24+ population. (PDF 24 kb)

Supplementary Figure 2

Characterization of three-dimensional structures produced in Matrigel cultures of CD29lo and CD29hi cells sorted from the Lin-CD24+ gate. (PDF 30 kb)

Supplementary Figure 3

Transplantation of mixed Rosa-26 and wild-type Lin-CD29hiCD24+ cells. (PDF 150 kb)

Supplementary Tables

This file contains Supplementary Tables 1–7. (DOC 78 kb)

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Shackleton, M., Vaillant, F., Simpson, K. et al. Generation of a functional mammary gland from a single stem cell. Nature 439, 84–88 (2006). https://doi.org/10.1038/nature04372

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