TY  - JOUR
TI  - Mammosphere Formation Assay Optimization in the Characterization of Cancer Stem Cells of the Primary Breast Tumor
AB  - Introduction&Objective: Breast cancer is the most frequently diagnosed cancer and the leading cause of death from cancer among females worldwide. Breast cancer tumors that feature breast cancer stem cells (BCSCs) are known to cause drug resistance and metastasis. Culturing BCSCs from primary tumors as mammospheres is both difficult and costly. This study aims to present an optimized mammosphere formation assay protocol and improve the breast cancer stem cell characterization process by determining the appropriate mammosphere forming method and proper cell density of cancer stem cells isolated from breast tumors. Material&Method: Cancer stem cells were isolated from breast tumors of two patients with invasive ductal carcinoma (IDC) and culturated. Subsequently, performed specific BCSCs surface markers and ALDH analysis by flow cytometry. Two mammosphere forming methods, i.e., low attachment and agar-coated wells seeded in three different cell concentrations, were compared. Results: CD44+, CD24- and CD133+ antibody expressions showed that these cells could be tumor-initiating CSCs. ALDH assay results also indicated that these cells possessed stem cell features. In addition, the results of the mammosphere assay revealed that agar-coated wells at a concentration of 7000 cells/cm2 had more prominent floating features and viable spheres. Conclusion: The findings of this study supported the hypothesis that agar-coated culture plates in mammosphere culture would increase the mammosphere formation efficiency (MFE) value and revealed the importance of the number of cells in elucidating the nature of BCSCs.
AU  - Uslu, Gizem
AU  - unal halbutogullari, zehra seda
AU  - GACAR, GÜLÇİN
AU  - Yazir, Yusufhan
AU  - Utkan, Nihat Zafer
DO  - 10.31067/acusaglik.1225228
PY  - 2023
JO  - Acıbadem Üniversitesi Sağlık Bilimleri Dergisi
VL  - 14
IS  - 2
SN  - 1309-470X
SP  - 108
EP  - 118
DB  - TRDizin
UR  - http://search/yayin/detay/1178219
ER  -