Unlocking the Potential of 3D Spheroid Cultures in Breast Cancer Stem Cell Enrichment and Isolation
Anan A. Ishtiah1, Omar Nafiis Hairuddin1, Badrul Hisham Yahaya21Department of Biomedical Sciences, Advanced Medical and Dental Institute (IPPT), University Sains Malaysia, Bertam 13200 Kepala Batas Penang, Malaysia, 2Breast Cancer Translational Research Program, Advanced Medical and Dental Institute (IPPT), University Sains Malaysia, Bertam Kepala Batas Penang, Malaysia,
Objectives: The present research conducted a comparative analysis of the efficacy of three-dimensional (3D) cultur ing methods in enriching and isolating breast cancer stem cells (BCSCs). The study compared multicellular spheroids grown in Matrigel and in suspension with the commonly used two-dimensional (2D) monolayer culturing method. Methods: The experiment involved a 9-day 3D multicellular spheroid culture, followed by a 24-hour monolayer culture using two breast cancer cell lines, namely MCF7 and MDA-MB-231. To evaluate BCSCs, the study assessed the expres sion of various surface markers, including CD44/CD24, Vimentin, and ALDH1, along with pluripotent stem cell genes like SOX2, OCT4, KLF4, and Nanog. Additionally, the Doxorubicin resistance and the capacity of single cells derived from each method to form spheroids in serum-free suspension culture were measured. Results: The findings revealed that 3D-cultured multicellular spheroids grown in suspension exhibited a significant increase in stem cell markers and Doxorubicin resistance. Furthermore, these spheroids demonstrated a higher ability to form single-cell spheroids with a size of more than 50 µm in a serum-free medium. Conclusion: Overall, this method of 3D culturing in suspension resulted in a substantial enrichment of BCSCs with enhanced self-renewal and proliferation capabilities when compared to both the 2D monolayer and 3D Matrigel meth ods. Consequently, this approach can serve as a crucial preliminary step in isolating BCSCs from cell lines using any available BCSCs isolation method. Keywords: Breast cancer, cancer resistance, cancer stem cells, doxorubicin, 3D culture
Cite This Article
Ishtiah A, Hairuddin O, Yahaya B. Unlocking the Potential of 3D Spheroid Cultures in Breast Cancer Stem Cell Enrichment and Isolation. EJMO. 2024; 8(3): 322-335
Corresponding Author: Badrul Hisham Yahaya