MCF7-CSCs came from the human breast cancer cell line MCF7 that was induced as previously described (38 (link)). Serum-free medium (SFM) consisted of DMEM/F12, 20 ng/mL basic fibroblast growth factor (Proteintech, Wuhan, China), 10 ng/mL epidermal growth factor (Proteintech), and 1×B27 supplement (Gibco Life Technologies, Lofer, Austria). When MCF7 cell density reached 90%, the cell precipitate was rinsed with phosphate-buffered saline (PBS), centrifuged at 1,000 rpm for 5 minutes, and the supernatant was discarded. MCF7 cells were resuspended in SFM, and the cell suspension was transferred to non-adherent culture flasks.
Protocol detail
Isolation of Breast Cancer Stem Cells
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Corresponding Organization : Guangdong Provincial Hospital of Traditional Chinese Medicine
Other organizations : Universidade Federal de São Carlos, Medway School of Pharmacy, University of Kent, King's College London, Medway NHS Foundation Trust, The Ohio State University Wexner Medical Center
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Variable analysis
independent variables
- Induction of MCF7 cells to become MCF7-CSCs
- Properties and characteristics of MCF7-CSCs
- Cell density of MCF7 cells (90%)
- Rinsing MCF7 cells with phosphate-buffered saline (PBS)
- Centrifugation of MCF7 cells at 1,000 rpm for 5 minutes
- Composition of serum-free medium (DMEM/F12, 20 ng/mL basic fibroblast growth factor, 10 ng/mL epidermal growth factor, and 1×B27 supplement)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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