Breast Cancer Stem Cell Characterization

Cells were stained with Aldefluor reagent as per manufacturer's instructions (Stemcell Technologies, Cat# 01700) and the percentage of cells with high Aldehyde Dehydrogenase (ALDH) activity were analyzed. Human breast cancer stem cell marker staining was determined by labeling human cells with CD44-APC and CD24-PE antibodies (BD Biosciences, Cat# 559942 and 555428). Dissociated mammary tumor cells and murine cell lines were stained with CD24-PE, CD29-FITC, CD31-APC, TER-119-APC (BD Biosciences, Cat# 553262, 561796, 561814, and 561033) and CD45-APC (Biolegend, Cat# 103111) antibodies, analyzed using the FACS Diva software, and sorted using the Aria Illu sorter (BD Biosciences) [47 (link)].

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Ruiz-Torres S.J., Benight N.M., Karns R.A., Lower E.E., Guan J.L, & Waltz S.E. (2017). HGFL-mediated RON signaling supports breast cancer stem cell phenotypes via activation of non-canonical β-catenin signaling. Oncotarget, 8(35), 58918-58933.

Publication 2017

Aldefluor reagent CD44-APC CD24-PE CD29-FITC CD31-APC TER-119-APC CD45-APC FACS Diva software

Aldehyde dehydrogenase Antibodies Aria Breast Cancer stem cell Cd44 Cell lines Cells Fitc Human Mammary tumor Murine Stemcell

Corresponding Organization : Veterans Health Administration

Other organizations : Cincinnati Children's Hospital Medical Center, University of Cincinnati

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Variable analysis

independent variables

Aldefluor reagent staining
CD44-APC and CD24-PE antibody labeling
CD24-PE, CD29-FITC, CD31-APC, TER-119-APC, and CD45-APC antibody staining

dependent variables

Percentage of cells with high Aldehyde Dehydrogenase (ALDH) activity
Human breast cancer stem cell marker staining

control variables

Manufacturer's instructions for Aldefluor reagent staining (Stemcell Technologies, Cat# 01700)
Antibody specifications (BD Biosciences, Cat# 559942, 555428, 553262, 561796, 561814, and 561033; Biolegend, Cat# 103111)
FACS Diva software for analysis
Aria Illu sorter (BD Biosciences) for sorting

controls

Not explicitly mentioned
Not explicitly mentioned

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