Molecular hierarchy of mammary differentiation yields refined markers of mammary stem cells

Dos Santos, C. O., Rebbeck, C., Rozhkova, E., Valentine, A., Samuels, A., Kadiri, L. R., Osten, P., Harris, E. Y., Uren, P. J., Smith, A. D., Hannon, G. J. (2013) Molecular hierarchy of mammary differentiation yields refined markers of mammary stem cells. Proceedings of the National Academy of Sciences of the United States of America, 110 (18). pp. 7123-7130. ISSN 0027-8424

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URL: http://www.ncbi.nlm.nih.gov/pubmed/23580620

DOI: 10.1073/pnas.1303919110

Abstract

The partial purification of mouse mammary gland stem cells (MaSCs) using combinatorial cell surface markers (Lin-CD24+CD29hCD49fh) has improved our understanding of their role in normal development and breast tumorigenesis. Despite the significant improvement in MaSC enrichment, there is presently no methodology that adequately isolates pure MaSCs. Seeking new markers of MaSCs, we characterized the stem-like properties and expression signature of label-retaining cells from the mammary gland of mice expressing a controllable H2b-GFP transgene. In this system, the transgene expression can be repressed in a doxycycline-dependent fashion, allowing isolation of slowly dividing cells with retained nuclear GFP signal. Here, we show that H2b-GFPh cells reside within the predicted MaSC compartment and display greater mammary reconstitution unit frequency compared with H2b-GFPneg MaSCs. According to their transcriptome profile, H2b-GFPh MaSCs are enriched for pathways thought to play important roles in adult stem cells. We found Cd1d, a glycoprotein expressed on the surface of antigen-presenting cells, to be highly expressed by H2b-GFPh MaSCs, and isolation of Cd1d+ MaSCs further improved the mammary reconstitution unit enrichment frequency to nearly a single-cell level. Additionally, we functionally characterized a set of MaSC-enriched genes, discovering factors controlling MaSC survival. Collectively, our data provide tools for isolating a more precisely defined population of MaSCs and point to potentially critical factors for MaSC maintenance.

Item Type:	Paper
Subjects:	Investigative techniques and equipment organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > differentiation Investigative techniques and equipment > flow cytometry organs, tissues, organelles, cell types and functions > tissues types and functions > mammary gland organs, tissues, organelles, cell types and functions organs, tissues, organelles, cell types and functions > cell types and functions > cell types > stem cells organs, tissues, organelles, cell types and functions > cell types and functions > cell types > stem cells organs, tissues, organelles, cell types and functions > cell types and functions > cell types > stem cells organs, tissues, organelles, cell types and functions > tissues types and functions
CSHL Authors:	Dos Santos, Camila O. Kadiri, Lolahon R. Osten, Pavel Rebbeck, Clare A. Rozhkova, Elena Samuels, Abigail Valentine, Amy Hannon, Gregory J.
Communities:	CSHL Cancer Center Program > Cancer Genetics CSHL Post Doctoral Fellows CSHL labs > Hannon lab CSHL labs > Osten lab CSHL Cancer Center Shared Resources > Flow Cytometry Service
Depositing User:	Matt Covey
Date:	2013
Date Deposited:	22 May 2013 18:30
Last Modified:	22 Dec 2017 17:11
PMCID:	PMC3645536
Related URLs:	Publisher
URI:	https://repository.cshl.edu/id/eprint/28305

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