SEARCH

SEARCH BY CITATION

Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
STEM_1001_sm_supplFig1.pdf155KSupplementary Figure 1. Fluorescence with BAA is sensitive to DEAB in all mammary fractions Plot shows MFI with BAA in the presence of DEAB divided by the MFI with BAA in the absence of DEAB, showing mean±SEM of this ratio for each mammary fractions (7 previously cryopreserved samples analyzed). Fractions are defined by the gating approach depicted in Fig. 1A.
STEM_1001_sm_supplFig2.pdf330KSupplementary Figure 2. Comparison of fluorescence with BAA in basal and primitive luminal fractions using CD10 as an additional phenotypic marker (A) Representative FACS profiles. Previously cryopreserved mammoplasty tissue was dissociated to form suspensions of single cells, incubated with Aldefluor in the presence or absence of DEAB, then stained with antibodies against CD49f, EpCAM, CD10, CD31 and CD45. Two fractions were identified based on relative expression of EpCAM and CD10, after gating on CD49f+CD31−CD45− cells; an EpCAM−/lowCD10+CD49f+CD31−CD45− and an EpCAM+CD10−CD49f+CD31−CD45− fraction, previously shown to be enriched for basal cells and primitive luminal cells respectively9. Lower panel displays the fluorescence with BAA in the 2 fractions in the presence (solid grey) or absence (open profiles) of DEAB. (B) Quantitation of the relative fluorescence intensity with BAA in the two fractions. Plot shows the MFI with BAA in the EpCAM+CD10−CD49f+CD31−CD45− fraction divided by the MFI with BAA in the EpCAM−/lowCD10+CD49f+CD31−CD45− fraction, in the absence of DEAB (mean±SEM of this ratio for 4 previously cryopreserved samples). The former (luminal progenitor enriched) exhibited a MFI that was 17±6 fold higher than the latter (basal) fraction. (C) Plot shows the MFI with BAA in the presence of DEAB divided by the MFI with BAA in the absence of DEAB (mean±SEM of this ratio for the two fractions for 4 previously cryopreserved samples).
STEM_1001_sm_supplFig3.pdf314KSupplementary Figure 3. Relative transcripts levels of all ALDH gene family members in different subsets of normal human mammary cells (A) Comparison of ALDH gene expression levels in 2 previously published whole transcriptome analyses of non-cultured primary human mammoplasty tissue samples. Upper panel: expression levels from array hybridization dataset3 for different fractions of cells isolated by FACS based on the expression of CD49f and EpCAM. Values plotted are the normalized (limma) signal intensity for each gene in each fraction divided by the average normalized (limma) signal intensity of a group of 7 housekeeping genes, averaged across all of the biological samples in the dataset. Lower panel: expression levels derived from LongSAGE dataset8 for different fractions of cells isolated immunomagnetically based on the expression of CD24, Muc1, CD44 and CD10. Values plotted are the tag counts for each gene in each fraction divided by the average tag count of a group of 7 housekeeping genes, averaged across all of the biological samples in the dataset. (B) Comparison of ALDH gene expression in previously published whole transcriptome array hybridization analysis of primary human mammoplasty tissue samples cells following 3 days adherent culture7. Different fractions of cells were subsequently isolated by FACS based on the expression of CD49f and EpCAM. Values plotted are the normalized (GC-RMA) signal intensity for each gene in each fraction divided by the average normalized signal intensity of a group of 7 housekeeping genes, averaged across all of the biological samples in the dataset.
STEM_1001_sm_supplMethods.pdf35KSupplementary Data
STEM_1001_sm_supplTables1-2.pdf54KSupplementary Tables 1 and 2. Results of 2° CFC assays used as regenerative endpoints for MRU assays

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.