Mammosphere Formation Assay from Human Breast Cancer Tissues and Cell Lines
Open Access
- 22 March 2015
- journal article
- research article
- Published by MyJove Corporation in Journal of Visualized Experiments
- No. 97,p. e52671
- https://doi.org/10.3791/52671
Abstract
Similar to healthy tissues, many blood and solid malignancies are now thought to be organised hierarchically, with a subset of stem-like cancer cells that self-renew while giving rise to more differentiated progeny. Understanding and targeting these cancer stem cells in breast cancer, which may possess enhanced chemo- and radio-resistance compared to the non-stem tumor bulk, has become an important research area. Markers including CD44, CD24, and ALDH activity can be assessed using fluorescence activated cell sorting (FACS) to prospectively isolate cells that display enhanced tumorigenicity when implanted into immunocompromised mice: the mammosphere assay has also become widely used for its ability to retrospectively identify sphere-forming cells that develop from single stem cell-like clones. Here we outline approaches for the appropriate culturing of mammospheres from cell lines or primary patient samples, their passaging, and calculations to estimate sphere forming efficiency (SFE). First we discuss key considerations and pitfalls in the appropriate planning and interpretation of mammosphere experiments.This publication has 23 references indexed in Scilit:
- Evolution of the Cancer Stem Cell ModelCell Stem Cell, 2014
- Mammosphere Formation in Breast Carcinoma Cell Lines Depends upon Expression of E-cadherinPLOS ONE, 2013
- Lineage Tracing Reveals Lgr5 + Stem Cell Activity in Mouse Intestinal AdenomasScience, 2012
- A restricted cell population propagates glioblastoma growth after chemotherapyNature, 2012
- Differentiation of breast cancer stem cells by knockdown of CD44: promising differentiation therapyJournal of Translational Medicine, 2011
- Eyes Wide Open: A Critical Review of Sphere-Formation as an Assay for Stem CellsCell Stem Cell, 2011
- Inducible formation of breast cancer stem cells and their dynamic equilibrium with non-stem cancer cells via IL6 secretionProceedings of the National Academy of Sciences of the United States of America, 2011
- let-7 Regulates Self Renewal and Tumorigenicity of Breast Cancer CellsCell, 2007
- CD44+/CD24-breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasisBreast Cancer Research, 2006
- Isolation and In vitro Propagation of Tumorigenic Breast Cancer Cells with Stem/Progenitor Cell PropertiesCancer Research, 2005