This paper describes the development of the Safety Performance Indicators (SPIs), the methodology for assessment of the safety culture of radiotherapy institutions using SPIs and common strengths and common areas for improvement. SPIs were categorized into eight sections which all together contain 23 attributes and each attribute has scoring criteria from 0 to 2 (in steps of 0.5). The maximum absolute cumulative score of SPIs was 46. A relative cumulative SPIs score of >80% indicates an institution strong commitment towards safety while score <50% indicates need for additional guidance to enhance safety culture. The assessment using SPIs was conducted for 17 radiotherapy institutions. The methodology of assessment includes interactive discussion, direct observations and document analysis. The relative cumulative SPIs score of seven institutions was found to be >80% while it was found in the range of 67.0% to 80% for the remaining ten institutions. Institutions were communicated about the cumulative SPIs score, areas of strengths, and areas for improvement. SPIs were found to be a good tool for safety culture assessment and can be utilized by the radiotherapy institutes for self-assessment to identify the areas of improvement. Based on SPIs score, regulatory body can grade the institutions from a radiation safety compliance point of view.