Related Experiment Video
Updated: May 23, 2025
Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors
Published on: March 1, 2020
Diversity of ER-positive and HER2-negative breast cancer stem cells attained using selective culture techniques
Satoshi Sueoka1,2, Azusa Kai1, Yukino Kobayashi3
1Department of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan.
Abstract:
Breast cancer stem cells are a promising therapeutic target in cancer. We explored breast cancer stem cell diversity and establish a methodology for selectively culturing breast cancer stem cells. We collected breast cancer tissues from surgical samples of treatment-naïve patients with estrogen receptor (ER)-positive, human epidermal growth factor receptor 2 (HER2)-negative breast cancer. Following isolation, cells were subjected to spheroid culture on non-adherent plates. Of the 57 cases, successful culture was achieved in 48 cases, among which the average ratio of CD44+/CD24- breast cancer cells increased from 13.8% in primary tumors to 61.6% in spheroids. A modest number of spheroid cells successfully engrafted in mice and subsequently re-differentiated within the murine environment, confirming their stemness. ER expression in spheroid cells exhibited negative conversion in 52.1% of cases. The proportion of Twist-, Snail-, and Vimentin-positive cells increased from 43.8%, 12.9%, and 7.7-75.0%, 58.1%, and 37.7%, respectively. ER-positive, HER2-negative breast cancer stem cells were classified into two groups using DNA microarrays. Gene Ontology analysis unveiled higher expression of immune response-related genes in one group and protein binding-associated genes in the other. We demonstrated stable and selective culture of breast cancer stem cells from patient-derived breast cancer tissue using spheroid cultures.
Insights
Researchers developed a method to selectively culture breast cancer stem cells (BCSCs) from patient tissues. This technique enhances the understanding and potential targeting of these crucial cells in estrogen receptor-positive, HER2-negative breast cancer.
Area of Science:
- Oncology
- Cell Biology
- Cancer Stem Cell Research
Background:
- Breast cancer stem cells (BCSCs) are implicated in tumor recurrence and therapy resistance.
- Targeting BCSCs offers a promising strategy for improving breast cancer treatment outcomes.
Purpose of the Study:
- To explore the diversity of BCSCs in estrogen receptor (ER)-positive, human epidermal growth factor receptor 2 (HER2)-negative breast cancer.
- To establish a reliable methodology for the selective culture of BCSCs from patient-derived tissues.
Main Methods:
- Spheroid culture of cells isolated from treatment-naïve breast cancer tissues.
- Analysis of cell surface markers (CD44+/CD24-), stemness markers (Twist, Snail, Vimentin), and ER expression.
- In vivo engraftment studies in mice.
- DNA microarrays and Gene Ontology analysis for BCSC classification.
Main Results:
- Successful spheroid culture achieved in 48 out of 57 cases, significantly increasing the CD44+/CD24- cell ratio.
- Demonstrated stemness of cultured cells through in vivo engraftment and re-differentiation.
- Observed ER negative conversion in 52.1% of spheroid cells and increased expression of EMT markers (Twist, Snail, Vimentin).
- Classified ER+/HER2- BCSCs into two distinct groups based on gene expression, with one group showing higher immune response genes and the other protein binding genes.
Conclusions:
- Spheroid culture provides a stable and selective method for culturing BCSCs from patient samples.
- The study highlights BCSC heterogeneity and potential plasticity, including ER status and EMT marker expression.
- Identified distinct molecular subtypes of ER+/HER2- BCSCs, paving the way for targeted therapeutic strategies.
