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Updated: May 2, 2026

Mammosphere Formation Assay from Human Breast Cancer Tissues and Cell Lines
Published on: March 22, 2015
Protocols for MCF-7 derived mammosphere generation and anti-cancer drug testing
Francesco Davide Naso1, Venturina Stagni2, Flavie Strappazzon3
1Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Fondazione Santa Lucia, Rome, Italy.
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Cancer research is a rapidly expanding field, requiring new methods and techniques to investigate the complex biology of the disease and its characteristics. A pivotal aspect of this research is the evaluation of the effects and efficacy of therapies. Here, we describe a simple protocol to generate mammospheres from MCF-7 cells, a useful model of breast cancer stem cells to evaluate cell death after therapy treatment. This subpopulation of cells, found in all types of tumors, is critically involved in therapeutic response as well as in disease recurrence and relapse, and therefore requires particular attention in oncology. The use of mammospheres represents a valuable approach to obtain preliminary in vitro results that closely resemble in vivo responses, providing an effective means to assess the efficacy of both conventional and next-generation therapeutic agents on breast cancer cell death.

