Dube, PhumuzileMonchusi, Bernice ATakundwa, Mutsa MKenmogne, Vanelle LMalise, AustinThimiri Govindaraj, Deepak B2025-09-252025-09-252025-031064-37451940-6029https://doi.org/10.1007/7651_2025_605http://hdl.handle.net/10204/14400This protocol outlines a detailed method for performing drug sensitivity testing (DST) on HeLa cells, focusing on both single-drug and combination-drug screening to assess cell viability. DST is integral to cancer research and functional precision medicine, providing insight into individual drug responses and facilitating the optimization of drug combinations. The protocol includes preparing and maintaining HeLa cell cultures, seeding in 96-well plates, and performing single and combination drug treatments using a low-throughput screening approach. These drug treatments aim to evaluate therapeutic effectiveness, enhance understanding of synergistic interactions, and identify optimal combinations that could minimize toxicity while overcoming resistance. Data analysis uses open-source tools, including the BREEZE pipeline and Synergy Finder, allowing for precise analysis of cell viability and drug interactions. This protocol provides a robust, reproducible framework for DST in cancer research, applicable to other cell lines, patient samples and various drug types/classes. The critical role of DST in improving clinical treatment strategies through precise, scalable drug response analysis is demonstrated.AbstractenCombination Drug assayDrug Sensitivity TestingDSTHeLa CellsPersonalized MedicineArticleN/A