Human primary cells are useful pre-clinical models as they more closely mimic the physiology of cells in vivo than continuous cell lines. Here, we explored some of the applications of human primary cells for cytotoxicity assays and drug screening. Cancer is a leading cause of female mortality worldwide and gynecologic cancers have a low survival rate. Further, cytotoxicity is a common side effect of all anti-cancer drugs. We investigated the cytotoxic effects of three anti-cancer drugs (alone or in combination) on three types of normal reproductive cells in vitro. Primary human uterine fibroblasts, cervical epithelial cells, and vaginal epithelial cells along with cervical (SiHa) and vaginal (VK2/E6E7) epithelial cancer cell lines were treated for two days with topotecan, paclitaxel, cisplatin, or a combination of topotecan and cisplatin at concentrations of 0 µM, 0.1 µM, 1 µM, 10 µM, or 100 µM. Cytotoxicity of these chemotherapeutic drugs was assessed by using Reliablue™ Cell Viability Reagent.