Background: Health professional graduates require the capacity to work safely, both clinically and culturally, when delivering care to Indigenous peoples worldwide. In the Australian context, the Aboriginal and Torres Strait Islander Health Curriculum Framework (The Framework) provides guidance for health professional programs to integrate, teach and assess Aboriginal and Torres Strait Islander peoples' (First Peoples) health content. There is, however, a lack of validated tools that measure the development of students' cultural capabilities. Objectives: To validate the Cultural Capability Measurement Tool with a cohort of health professional students. Design: A descriptive cohort design was used. Setting and Participants: All students (N = 753) enrolled in a discrete First Peoples Health course at an Australian university were invited to complete the Cultural Capability Measurement Tool. Methods: The tool was tested for reliability, content and construct validity using confirmatory factor analysis; and concurrent validity using and the Cultural Understanding Self-Assessment Tool. Results: A sample of 418 (73% response rate) was recruited. Most participants were enrolled in the Bachelor of Nursing program (n = 369, 82%). The Cultural Capability Measurement Tool had a Cronbach's alpha coefficient of 0.86. A five-factor solution was confirmed which reflected the cultural capability domains and accounted for 51% of the variance. Scores correlated with students' cultural understanding (r = 0.28, p < 0.001). Conclusions: Successful implementation of The Framework requires instruments to measure changes in students' cultural capabilities. Measuring nursing students' cultural capabilities can inform their development, identify areas of strengths and deficits for educators, and will ultimately contribute to the development of a culturally safe nursing workforce.