Patient-Derived Primary Cultures in Breast Cancer: A Comparative Study of Isolation and Culture Methods

Introduction: Breast cancer is the most common cancer among women worldwide. Primary culture methods play a pivotal role in understanding cancer’s heterogeneity. Improving experimental conditions is essential for advancements in treatment. This study aims to compare two different isolation methods for primary culture derived from breast cancer patient tissues.
Methods: Breast tissues surgically excised from breast cancer patients were cultured under sterile conditions using explant culture and enzymatic digestion. In the explant culture group, parallel lines were drawn in the wells to examine cell adhesion, while the effect of collagen on cells was assessed by coating the wells and evaluating them on days 0, 5, and 10. Enzymatic digestion was analyzed using two different methods: short-term and long-term. The morphology of cultured cells was examined under an inverted microscope.
Results: Long-term enzymatic digestion resulted in better cell adhesion and proliferation compared to short-term, leading to higher cell counts and more fibroblast-like morphology. However, explant culture yielded the best results in terms of cell count and morphology for both tissues. The cells in explant cultures exhibited epithelioid morphology from day 5 onward, maintaining this characteristic through day 10. Additionally, no significant effects were observed on collagen-coated surfaces in our study.
Discussion and Conclusion: The findings highlight the importance of selecting appropriate primary culture methods in breast cancer research. While long-term enzymatic digestion improves proliferation, explant culture remains the superior approach for preserving cell morphology and mimicking the tumor microenvironment. Optimizing culture techniques is essential for enhancing translational cancer research and advancing therapies.

Keywords: Breast cancer; Explant culture; Patient derived primary culture