Abstract
Previous studies have demonstrated that normal mouse mammary tissue contains a rare subset of mammary stem cells. We now describe a method for detecting an analogous subpopulation in normal human mammary tissue. Dissociated cells are suspended with fibroblasts in collagen gels, which are then implanted under the kidney capsule of hormone-treated immunodeficient mice. After 2–8 weeks, the gels contain bilayered mammary epithelial structures, including luminal and myoepithelial cells, their in vitro clonogenic progenitors and cells that produce similar structures in secondary transplants. The regenerated clonogenic progenitors provide an objective indicator of input mammary stem cell activity and allow the frequency and phenotype of these human mammary stem cells to be determined by limiting-dilution analysis. This new assay procedure sets the stage for investigations of mechanisms regulating normal human mammary stem cells (and possibly stem cells in other tissues) and their relationship to human cancer stem cell populations.
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Acknowledgements
C3H 10T1/2 mouse embryonic fibroblasts were a kind gift from G. Cunha, University of California, San Francisco. The authors acknowledge the excellent technical contributions of D. Wilkinson, G. Edin, the staff of the Flow Cytometry Facility of the Terry Fox Laboratory and the Centre for Translational and Applied Genomics. Mammoplasty tissue was obtained with the assistance of J. Sproul, P. Lennox, N. Van Laeken and R. Warren. This project was funded by grants from Genome British Columbia and Genome Canada, the Canadian Stem Cell Network and the Canadian Breast Cancer Foundation British Columbia and Yukon Division. P.E. was a recipient of a US Department of Defense Breast Cancer Research Program Studentship, a Terry Fox Foundation Research Studentship from the National Cancer Institute of Canada, a Canadian Imperial Bank of Commerce interdisciplinary award and a Canadian Stem Cell Network Studentship. J.S. held a Canadian Breast Cancer Foundation British Columbia and Yukon Division Fellowship and a Canadian National Science and Engineering Research Council Industrial Fellowship. A.R. held a Canadian Breast Cancer Foundation British Columbia and Yukon Division Fellowship and a Canadian Institutes of Health Research Fellowship. G.T. holds a Canadian Institutes of Health Research Pathology Training Fellowship. S.A. is supported by a Canada Research Chair in Molecular Oncology. The Centre for Translational and Applied Genomics laboratory is supported by a Canadian Institutes for Health Research Resource award.
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P.E. designed and conducted most of the experiments and drafted the manuscript. J.S. initiated the work that led to the gel implant protocol, undertook preliminary experiments and contributed to the writing of the manuscript. A.R. critiqued the manuscript and participated in discussions of the experiments. G.T. and S.A. reviewed the histological preparations and contributed to the writing of the manuscript. J.T.E. helped organize the accrual of the mammoplasty material used. C.J.E. conceptualized the study and finalized the writing of the manuscript.
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Eirew, P., Stingl, J., Raouf, A. et al. A method for quantifying normal human mammary epithelial stem cells with in vivo regenerative ability. Nat Med 14, 1384–1389 (2008). https://doi.org/10.1038/nm.1791
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DOI: https://doi.org/10.1038/nm.1791
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