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Robotic Mammosphere Assay for High-Throughput Screening in Triple-Negative Breast Cancer

Journal article
Authors Paul A. Fitzpatrick
Nina Akrap
Elin Söderberg
H. Harrison
G. J. Thomson
Göran Landberg
Published in Slas Discovery
Volume 22
Issue 7
Pages 827-836
ISSN 2472-5552
Publication year 2017
Published at Sahlgrenska Cancer Center
Pages 827-836
Language en
Keywords cancer and cancer drugs, high-content screening, stem cells, cell-based assays, stem-cells, identification, resistance, inhibitor, target, Biochemistry & Molecular Biology, Biotechnology & Applied Microbiology, Chemistry
Subject categories Cancer and Oncology


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.

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