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Since the discovery that neural tissue contains a population of stem cells that form neurospheres in vitro, sphere-forming assays have been adapted for use with a number of different tissue types for the quantification of stem cell activity and self-renewal. One tissue type widely used for stem cell investigations is mammary tissue, and the mammosphere assay has been used in both normal tissue and cancer. Although it is a relatively simple assay to learn, it can be difficult to master. There are methodological and analytical aspects to the assay which require careful consideration when interpreting the results. We describe here a detailed mammosphere assay protocol for the assessment of stem cell activity and self-renewal, and discuss how data generated by the assay can be analysed and interpreted.

Original publication




Journal article


J mammary gland biol neoplasia

Publication Date





111 - 117


Animals, Breast Neoplasms, Carcinoma, Intraductal, Noninfiltrating, Cell Line, Cell Line, Tumor, Cell Proliferation, Female, Humans, Mammary Glands, Animal, Mammary Glands, Human, Neoplastic Stem Cells, Tumor Stem Cell Assay