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Updated: May 16, 2026
Isolation and Functional Assessment of Human Breast Cancer Stem Cells from Cell and Tissue Samples
Published on: October 2, 2020
[Isolation and characterization of human breast tumor stem cells]
Li-bin Wang1, Ya-qin He, Li-gang Wu
1Institute of Stem Cells, General Hospital, Ministry of Education, Ningxia Medical University, Yinchuan 750004, China. wanglinbin007@126.com
Aim:
To isolate and culture human breast cancer stem cells, and characterize their biological properties in vitro.
Methods:
Fresh tumor tissues of breast cancer patients were collected from surgical rooms. The tissue samples were mechanically sheared and enzyme-digested to get single cell suspensions. Cancer cell surface antigens were analyzed by flow cytometry, and cell growth curve was established by MTT assay; Real-time quantitative PCR (qPCR) was employed to detect the expressions of stem cell-specific genes Sox2 and Nanog in the cancer cells; Immunofluorescence staining was performed to examine the expressions of breast cancer-specific proteins Bcl-2 and progesterone receptor (PR).
Results:
Human breast cancer stem cells grew in the form of spheres. These stem cells had a CD44(+);/CD24(-); phenotype as characterized by flow cytometry, expressed high levels of Sox2 and Nanog genes, and were positive for Bcl-2 and PR.
Conclusion:
Human breast tumors contain CD44(+);/CD24(-); cancer stem cells which are capable of self-renewal and proliferation, and can be long-term cultured and expanded in vitro.
