Background Vaccine hesitancy remains a significant issue threatening public health. The study aimed to determine the vaccination status of patients undergoing hemodialysis for chronic kidney disease and to analyze their levels of vaccine hesitancy. Methods This cross-sectional analytical study was conducted in hemodialysis centers located in Ankara, the capital of Turkey. Data collection involved a structured questionnaire capturing sociodemographic details, comorbidities, vaccination history, and hesitancy levels, coupled with the Vaccine Hesitancy Scale (VHS), a 9-item tool validated in Turkish that measures trust in vaccines and perceived risks. Results Among the 548 participants (mean age: 60.4 +/- 12.9 years, range: 18-93; 57.8% male), 38.9% had a high school education or higher, and 52.7% reported income below their expenses. Despite 92.7% having received at least one vaccine during adulthood, knowledge about vaccines was limited, with only 7.7% answering all vaccine-related questions correctly. Influenza (73.4%) and hepatitis B (58.4%) were the most administered vaccines. The median VHS score was 32 (range: 11-45), with 84.7% scoring above 25, reflecting low overall hesitancy. Participants who identified physicians (76.6%) and healthcare workers (57.5%) as trusted sources of vaccine information tended to have lower hesitancy scores (p < 0.001). No significant associations were found between hesitancy and gender, education, income level, or general health perception. Conclusions While vaccination rates among hemodialysis patients are relatively high, critical gaps in knowledge persist, emphasizing the need for targeted educational programs. The active involvement of healthcare professionals is crucial to reduce hesitancy and enhance vaccine confidence in this vulnerable population.