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Background  Accumulating evidence supports cancer to initiate and develop from a small population of stem-like cells termed as cancer stem cells (CSC). The exact phenotype of CSC and their counterparts in normal mammary gland is not well characterized. In this study our aim was to evaluate the phenotype and function of stem/progenitor cells in normal mammary epithelial cell populations and their malignant counterparts.    Methods  Freshly isolated cells from both normal and malignant human breasts were sorted using 13 widely used stem/progenitor cell markers individually or in combination by multi-parametric (up to 9 colors) cell sorting. The sorted populations were functionally evaluated by their ability to form colonies and mammospheres,  in vitro .    Results  We have compared, for the first time, the stem/progenitor markers of normal and malignant breasts side-by-side. Amongst all markers tested, we found CD44 high /CD24 low  cell surface marker combination to be the most efficient at selecting normal epithelial progenitors. Further fractionation of CD44 high /CD24 low  positive cells showed that this phenotype selects for luminal progenitors within Ep-CAM high /CD49f + cells, and enriches for basal progenitors within Ep-CAM -/low /CD49f + cells. On the other hand, primary breast cancer samples, which were mainly luminal Ep-CAM high , had CD44 high /CD24 low  cells among both CD49f neg  and CD49f + cancer cell fractions. However, functionally, CSC were predominantly CD49f + proposing the use of CD44 high /CD24 low  in combination with Ep-CAM/CD49f cell surface markers to further enrich for CSC.    Conclusion  Our study clearly demonstrates that both normal and malignant breast cells with the CD44 high /CD24 low  phenotype have the highest stem/progenitor cell ability when used in combination with Ep-CAM/CD49f reference markers. We believe that this extensive characterization study will help in understanding breast cancer carcinogenesis, heterogeneity and drug resistance.

bmc cancer

Profiling of normal and malignant breast tissue show CD44high/CD24lowphenotype as a predominant stem/progenitor marker when used in combination with Ep-CAM/CD49f markers