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Updated: Mar 5, 2026
Mammosphere Formation Assay from Human Breast Cancer Tissues and Cell Lines
Published on: March 22, 2015
Robotic Mammosphere Assay for High-Throughput Screening in Triple-Negative Breast Cancer
P A Fitzpatrick1, N Akrap1, E M V Söderberg1
11 Sahlgrenska Cancer Center, Göteborgs Universitet, Göteborg, Sweden.
Abstract:
In order to identify novel treatment principles specifically affecting cancer stem cells in triple-negative breast cancer, we have developed a high-throughput screening method based on the mammosphere and anoikis resistance assays allowing us to screen compounds using a functional readout. The assay was validated against manual protocols and through the use of positive controls, such as the response to hypoxia and treatment with the known cancer stem cell-targeting compound salinomycin. Manual and robotic procedures were compared and produced similar results in cell handling, cell cultures, and counting techniques, with no statistically significant difference produced from either method. The variance between samples processed manually versus robotically was no greater than 0.012, while Levene's test of significance was 0.2, indicating no significant difference between mammosphere data produced manually or robotically. Through the screening of 989 FDA-approved drugs and a follow-up screen assessing the antineoplastic subgroup, we have identified three therapeutic compounds with the ability to modulate the breast cancer stem cell fraction in the triple-negative breast cancer cell line MDA-MB-231, highlighting their potential usage as stem cell-specific adjuvant treatments.
Insights
Researchers identified novel treatments for triple-negative breast cancer (TNBC) by screening FDA-approved drugs. Three compounds effectively targeted cancer stem cells, offering potential as new adjuvant therapies.
Area of Science:
- Oncology
- Biotechnology
- Pharmacology
Background:
- Triple-negative breast cancer (TNBC) is aggressive and lacks targeted therapies.
- Cancer stem cells (CSCs) drive tumor recurrence and metastasis in TNBC.
- Novel therapeutic strategies targeting CSCs are urgently needed.
Purpose of the Study:
- To develop and validate a high-throughput screening (HTS) assay for identifying CSC-targeting agents in TNBC.
- To screen FDA-approved drugs for their ability to modulate the CSC fraction in TNBC.
- To identify potential novel adjuvant treatments for TNBC.
Main Methods:
- Development of an HTS assay using mammosphere and anoikis resistance assays for functional readout.
- Validation of the HTS assay against manual protocols and with positive controls (hypoxia, salinomycin).
- Comparison of manual versus robotic procedures, showing no significant difference in results.
- Screening of 989 FDA-approved drugs, followed by a focused screen of the antineoplastic subgroup.
Main Results:
- The HTS assay demonstrated reliability and reproducibility, comparable between manual and robotic methods.
- Three compounds were identified that significantly modulate the cancer stem cell fraction in MDA-MB-231 TNBC cells.
- These compounds showed potential as stem cell-specific agents.
Conclusions:
- A validated HTS platform can effectively identify CSC-targeting compounds in TNBC.
- Specific FDA-approved drugs show promise as novel adjuvant therapies by targeting TNBC CSCs.
- Further investigation into these compounds could lead to improved treatment strategies for TNBC.
