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

Abstract

BackgroundAccumulating 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.MethodsFreshly 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.ResultsWe have compared, for the first time, the stem/progenitor markers of normal and malignant breasts side-by-side. Amongst all markers tested, we found CD44high/CD24low cell surface marker combination to be the most efficient at selecting normal epithelial progenitors. Further fractionation of CD44high/CD24low positive cells showed that this phenotype selects for luminal progenitors within Ep-CAMhigh/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-CAMhigh, had CD44high/CD24low cells among both CD49fneg and CD49f + cancer cell fractions. However, functionally, CSC were predominantly CD49f + proposing the use of CD44high/CD24low in combination with Ep-CAM/CD49f cell surface markers to further enrich for CSC.ConclusionOur study clearly demonstrates that both normal and malignant breast cells with the CD44high/CD24low 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.

Optimization of culture medium for the isolation and propagation of human breast cancer cells from primary tumour biopsies

“…In a previous study by Al-Hajj et al (2003), primary cells contained a sub-population of CD44 + CD24 -cells with high tumourigenicity (Al-Hajj et al, 2003). However, the possibility that the primary cells expressing CD44 + CD24 -were not all BCSCs appears likely (Ghebeh et al, 2013;Mannello, 2013). In a recent study, Ghebeh et al (2013) clearly demonstrated that both normal breast tissue and breast cancer tissue harboured CD44 + CD24 -cells (Ghebeh et al, 2013).…”

“…In a recent study, Ghebeh et al (2013) clearly demonstrated that both normal breast tissue and breast cancer tissue harboured CD44 + CD24 -cells (Ghebeh et al, 2013). They also suggested that BCSCs would be enriched in the CD44 + CD24 -cells if they were combined with the CD49f + phenotype (Ghebeh et al, 2013). CD49f was also determined as a marker of BCSCs in previous studies (Meyer et al, 2010;Yu et al, 2012 However, Medium D and Medium M also supported stromal cell proliferation.…”

“…However, the possibility that the primary cells expressing CD44 + CD24 -were not all BCSCs appears likely (Ghebeh et al, 2013;Mannello, 2013). In a recent study, Ghebeh et al (2013) clearly demonstrated that both normal breast tissue and breast cancer tissue harboured CD44 + CD24 -cells (Ghebeh et al, 2013). They also suggested that BCSCs would be enriched in the CD44 + CD24 -cells if they were combined with the CD49f + phenotype (Ghebeh et al, 2013).…”

Obesity and p16INK4A Downregulation Activate Breast Adipocytes and Promote Their Protumorigenicity

“…Normal breast luminal cells were obtained from the same tissues and were cultured as previously described (51).…”

Impact of Progesterone on Stem/Progenitor Cells in the Human Breast

“…While they do not initiate tumor formation, they have been postulated to drive the growth of the tumor mass [23]. It has been suggested that CSCs may derive from normal progenitor counterparts, because the CD44+/CD24-markers [24] which, with the aldehyde dehydrogenase enzyme (ALDH) [25] currently define CSCs, have been used to isolate cells with progenitor activity from normal human breast epithelial cells [26]. These observations highlight the fact that tight regulation of cell fate determination is critical for normal breast development, and that disrupted cell fate decisions are implicated in breast cancer.…”