Background: Vaccines are medical products with a short shelf life and are easily damaged by deviations in temperature from the recommended ranges. Vaccines lose their quality if the cold chain system is not properly managed. Cold chain management is still a major challenge in developing countries, including India. Thus, this study aimed to assess vaccine cold chain management and associated factors at public health facilities. Methods: A facility-based cross-sectional study design was applied. Data was collected using the observation check list and interviewer- administered pre-tested structured questionnaires.54 cold chain handlers of 27 facilities of district Gwalior were interviewed Multivariate analysis at a p-value <0.05 and OR with 95% CI was used to measure the degree of association between independent variables and the outcome variable. Results: The study indicates that 43 (79.6%) public health facilities had good/average cold chain management practice. Experience greater than 2 years (OR=5.1, 95%CI=1.2;22.0), cold chain guidelines (OR=7.59 (95%,CI=1.4;39.6)), training on cold chain management (OR=8.40,95%CI=1.4–9.8), and supportive supervision on cold chain management (4.50 (95% CI=1.0;19.4)) were statistically significantly associated with good cold chain management practice. Conclusion: The result of the study indicated that there was not satisfactory cold chain management practice in the study area. Strengthening the knowledge of healthcare workers and supportive supervision on cold chain management by giving training and monitoring their practice toward cold chain management may help to improve the cold chain management practice.