Many health professions faculty members lack training on fundamental lesbian, gay, bisexual, transgender, and queer (LGBTQ+) health topics. Faculty development is needed to address knowledge gaps, improve teaching, and prepare students to competently care for the growing LGBTQ+ population. We conducted a program evaluation of the massive open online course Teaching LGBTQ+ Health: A Faculty Development Course for Health Professions Educators from the Stanford School of Medicine. Our goal was to understand participant demographics, impact, and ongoing maintenance needs to inform decisions about updating the course. We evaluated the course for the period from March 27, 2021, to February 24, 2023, guided by the RE-AIM (Reach, Effectiveness, Adoption, Implementation, and Maintenance) framework. We assessed impact using participation numbers, evidence of learning, and likelihood of practice change. Data included participant demographics, performance on a pre- and postcourse quiz, open-text entries throughout the course, continuing medical education (CME) credits awarded, and CME course evaluations. We analyzed demographics using descriptive statistics and pre- and postcourse quiz scores using a paired 2-tailed t test. We conducted a qualitative thematic analysis of open-text responses to prompts within the course and CME evaluation questions. Results were reported using the 5 framework domains. Regarding Reach, 1782 learners participated in the course, and 1516 (85.07%) accessed it through a main course website. Of the different types of participants, most were physicians (423/1516, 27.9%) and from outside the sponsoring institution and target audience (1452/1516, 95.78%). Regarding Effectiveness, the median change in test scores for the 38.1% (679/1782) of participants who completed both the pre- and postcourse tests was 3 out of 10 points, or a 30% improvement (P<.001). Themes identified from CME evaluations included LGBTQ+ health as a distinct domain, inclusivity in practices, and teaching LGBTQ+ health strategies. A minority of participants (237/1782, 13.3%) earned CME credits. Regarding Adoption, themes identified among responses to prompts in the course included LGBTQ+ health concepts and instructional strategies. Most participants strongly agreed with numerous positive statements about the course content, presentation, and likelihood of practice change. Regarding Implementation, the course cost US $57,000 to build and was intramurally funded through grants and subsidies. The course faculty spent an estimated 600 hours on the project, and educational technologists spent another 712 hours. Regarding Maintenance, much of the course is evergreen, and ongoing oversight and quality assurance require minimal faculty time. New content will likely include modules on transgender health and gender-affirming care. Teaching LGBTQ+ Health improved participants' knowledge of fundamental queer health topics. Overall participation has been modest to date. Most participants indicated an intention to change clinical or teaching practices. Maintenance costs are minimal. The web-based course will continue to be offered, and new content will likely be added.