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Objectives Training healthcare professionals in emergency maternal healthcare is a critical component of improving overall maternal health in developing countries like Rwanda. This paper explored the challenges that healthcare professionals who participated in a Continuing Professional Development (CPD) program on Advanced Life Support in Obstetrics® (ALSO) face in putting the learned knowledge and skills into practice in hospitals of Rwanda. Methods This study used a mixed methods approach to understand the challenges/barriers to applying new knowledge and skills in the hospitals of Rwanda. We conducted thirteen purposive in-depth interviews with ALSO® trainees (nurses, midwives and physicians) complemented with a cross-sectional survey on staff turnover in eight of the nine hospitals in the Eastern province of Rwanda. Results Our study found that trainees do not get enough opportunity to apply the new knowledge and skills in their hospitals and expand to health centers. In part because they are frequently rotating to different departments of the hospital and are not getting the opportunity to train their colleagues to share the learned knowledge and skills. The lack of refresher trainings/mentorship and the high personnel turnover were also reported as a barrier to applying new knowledge and skills. Reasons for staff turnover included pursuing further studies, a better opportunity (job/remuneration), low morale, and family related motives including joining a spouse or better schools for children. Conclusions for Practice Expanding and formalizing CPD training to all the healthcare professionals involved in providing maternal care services would improve the provision of emergency maternal healthcare in Rwanda. [ABSTRACT FROM AUTHOR]