Background and Objective: Cisplatin (CP) is a chemotherapy drug extensively utilized for its efficacy in treating a variety of cancer types. Despite its therapeutic benefits, CP is notably associated with renal toxicity as a side effect. The ongoing research into mitigating agents against CP-induced renal toxicity focuses on substances with antioxidant and anti-inflammatory characteristics. This investigation aims to explore the potential protective properties of a cell culture medium derived from human amniotic fluid (AMX) against renal toxicity prompted by CP in a rodent model. Materials and Methods: Fifty-five male Wistar albino rats were divided into five groups: Control, AMX, CP, CP+AMX and AMX+CP. All agents were administered intraperitoneally. The study assessed serum parameters, histopathological changes and oxidative/antioxidative markers in the kidneys. Blood urea nitrogen (BUN) and creatinine (Cre) levels were measured to evaluate renal function, while total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI) were analyzed for oxidative damage. Results: The AMX pre-treatment group showed a significant decrease in BUN and Cre levels, indicating a reduction in CP-induced renal damage. Histopathologically, AMX pre-treatment reduced inflammation, congestion and edema in the kidneys. Although TAS levels increased post-CP administration, no significant changes were observed in TOS and OSI values. The study demonstrates that AMX can mitigate CP-induced nephrotoxicity, as evidenced by improved renal function markers and reduced histopathological damage. Conclusion: The protective effects of AMX may be attributed to its anti-inflammatory properties and the presence of growth factors and glutamine. While promising, further research is required to fully elucidate AMX’s protective mechanisms against CP-induced renal damage.