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.
AIM: To isolate and culture humanbreast cancer stem cells, and characterize their biological properties in vitro. METHODS: Fresh tumor tissues of breast cancerpatients 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:Humanbreast 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:Humanbreast 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.
Authors: Deep Shah; Debra Wyatt; Andrew T Baker; Patricia Simms; Daniel S Peiffer; Michelle Fernandez; Emad Rakha; Andrew Green; Alexandra Filipovic; Lucio Miele; Clodia Osipo Journal: Clin Cancer Res Date: 2018-06-12 Impact factor: 12.531