My Vital Cycles® is a holistic ovulatory menstrual health literacy program for adolescent females. A whole school approach was used when trialled in one single-sex secondary school in Perth Australia. This study investigated the enablers and barriers for its future implementation from the perspective of the school's teachers, nurses and psychologists. The design was a pre-trial survey of ovulatory menstrual health knowledge and attitudes, followed by post-trial interviews and focus group discussions (FGDs). In the pre-trial survey, 12 school staff demonstrated positive attitudes to ovulatory menstrual health; sound anatomical knowledge; but limited knowledge of cycle patterns. There were two FGDs with five participants and eight post-trial semi-structured interviews, giving 13 school staff in total. Deductive thematic analysis identified 14 themes within a whole school approach framework, specifically: policies and resources; community partnerships; curriculum; social-emotional environment; and school health services. Enablers included school staff's positive attitudes; the program's validated materials; its curricular alignment; and address of stigma. Barriers included school staff's need for professional development; extending the program over years; Health and Science silos; dependence on school leadership; and challenges of building community partnerships. Co-design with school leadership and staff to enhance opportunities for a whole school approach is recommended for future implementation.