How to cite this article: Donnenberg VS, Donnenberg AD, Zimmerlin L, Landreneau RJ, Bhargava R, Wetzel RA, Basse P, Brufsky AM. Localization of CD44 and CD90 positive cells to the invasive front of breast tumors. Cytometry Part B 2010; 78B: 287–301.
Original Article
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Localization of CD44 and CD90 positive cells to the invasive front of breast tumors†‡§
Article first published online: 30 APR 2010
DOI: 10.1002/cyto.b.20530
Copyright © 2010 International Clinical Cytometry Society
Additional Information
How to Cite
Donnenberg, V. S., Donnenberg, A. D., Zimmerlin, L., Landreneau, R. J., Bhargava, R., Wetzel, R. A., Basse, P. and Brufsky, A. M. (2010), Localization of CD44 and CD90 positive cells to the invasive front of breast tumors. Cytometry, 78B: 287–301. doi: 10.1002/cyto.b.20530
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PE phycoerythrin, PECy5 phycoerythrin cyanine 5, SHH sonic hedgehog, SSc side light scatter, UCI upper confidence interval
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Vera S. Donnenberg, Ph.D., and Albert D. Donnenberg, Ph.D., codesigned the experiments, analyzed the data, and cowrote the manuscript. Ludovic Zimmerlin, M.S., developed, validated, and performed immunofluorescent staining. Rodney J. Landreneau, M.D., and Adam M. Brufsky, M.D., provided clinical specimens and interpretation of therapy and clinical course. Rohit Bargava, M.D., prepared the tissue microarrays and reviewed immunohistostaining and histology. Per Basse, M.D., Ph.D., supervised the tumorigenicity experiments. Ryan Wetzel validated and performed immunohistochemical staining.
Publication History
- Issue published online: 25 AUG 2010
- Article first published online: 30 APR 2010
- Manuscript Accepted: 2 APR 2010
- Manuscript Revised: 29 MAR 2010
- Manuscript Received: 26 AUG 2009
Funded by
- Department of Defense. Grant Numbers: BC032981, BC044784
- Hillman Foundation
- Glimmer of Hope Foundation
Keywords:
- cancer stem cells;
- breast cancer;
- flow cytometry;
- immunohistochemistry;
- immunofluorescence;
- cytokeratin;
- CD44;
- CD90;
- CD133;
- CD117
Abstract
Background:
A variety of markers have been proposed to identify breast cancer stem cells. Here, we used immunohistostaining and flow cytometry to analyze their interrelationships and to sort cells for tumorigenicity studies.
Methods:
Cytokeratin, CD44, and CD90 were localized to primary breast cancer and normal breast (NB) tissue by immunohistostaining and related to CD117 and CD133 expression by flow cytometry. Immunodeficient NOD.CB17-Prkdcscid/J and NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice were used to test tumorigenicity of sorted CD90+ low-light scatter, CD90+ high-light scatter, and CD90neg tumor cells.
Results:
NB basal cells coexpressed CD44 and CD90. As cells transited luminally, CD44 was retained and downmodulated, and CD90 was lost and cytokeratin increased. In breast tumors, basal-like CD44+/CD90+ cells were localized to the tumor periphery, adjacent to CD90+ stroma. Like normal luminal cells, interior tumor cells were CD44+/CD90−. Immunophenotyping (CD44/CD90/CD117/CD133) of cytokeratin+ cells revealed no significant difference in expression between tumors and tumor-free breast. In both, CD133 was distributed approximately equally among CD44/CD90 subsets, whereas CD117 expression was highest in the basal-associated CD44+/CD90+ subset. Sorted CD90+ pleural effusion cells with lymphoid light scatter, 49% of which were CD44+, were uniquely tumorigenic in immunodeficient mice (100 cells/injection).
Conclusions:
Our data demonstrate that all tumors contain a small population of CD44+/CD90+ cells, mimicking the phenotype of ductal-basal cells. These are localized to the tumor periphery, adjacent to CD90+ stroma. Among the nonhematopoietic, nonmesothelial cells found in metastatic pleural effusions, low-light scatter CD90+ cells are most potently tumorigenic, compared to high-scatter CD90+ cells and CD90− cells. © 2010 International Clinical Cytometry Society