Density Gradient Centrifugation is an Effective Tool to Isolate Cancer Stem-Like Cells from Hypoxic and Normoxia Triple-Negative Breast Cancer Models
Abstract
:1. Introduction
2. Results
2.1. Fractionation of Breast Cancer Cells by Density Gradient Centrifugation and CSCs’ Enriched Fraction Determination via Non-Adherent 3D Mammosphere Assay
2.2. Fractionation of Mammospheres by Size Using Density Gradient Centrifugation
2.3. CSCs from TNBC Have Hybrid Metabolism and CSCs from Large Mammospheres Have Increased Glycolytic Capacity
2.4. HYOU1 Is Highly Expressed in Hyoxic TNBC and HYOU1 Is Related with Aggressiveness, Stemness, Hyper-Metabolic Activity, Metastasis and Poor Prognosis of Breast Caner
3. Discussion
4. Materials and Methods
4.1. Experimental Model Cell Lines
4.2. Gradient-Based Breast CSCs and Hypoxic Mammosphere Separation
4.3. Three-Dimensional Mammosphere Growth and Three-Dimensional Mammosphere Formation Assay and Colony Formation Assay
4.4. Fluorescent Microscopy Analysis
4.5. ATP Assay Using Cell-Titer-Glo
4.6. Metabolic Flux Analysis for Density Gradient Separated Breast Cancer Cells
4.7. Viral Transduction of HYOU1 Gene
4.8. Bioinformatics Analysis
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Sargiacomo, C.; Klepinin, A. Density Gradient Centrifugation is an Effective Tool to Isolate Cancer Stem-Like Cells from Hypoxic and Normoxia Triple-Negative Breast Cancer Models. Int. J. Mol. Sci. 2024, 25, 8958. https://doi.org/10.3390/ijms25168958
Sargiacomo C, Klepinin A. Density Gradient Centrifugation is an Effective Tool to Isolate Cancer Stem-Like Cells from Hypoxic and Normoxia Triple-Negative Breast Cancer Models. International Journal of Molecular Sciences. 2024; 25(16):8958. https://doi.org/10.3390/ijms25168958
Chicago/Turabian StyleSargiacomo, Camillo, and Aleksandr Klepinin. 2024. "Density Gradient Centrifugation is an Effective Tool to Isolate Cancer Stem-Like Cells from Hypoxic and Normoxia Triple-Negative Breast Cancer Models" International Journal of Molecular Sciences 25, no. 16: 8958. https://doi.org/10.3390/ijms25168958