LGBTQ+ communities experience significant healthcare and academic success disparities due to barriers in care, negative attitudes towards them, and lack of awareness among healthcare providers and educators. Addressing the unique health and wellbeing concerns of LGBTQ+ individuals in the classroom and clinic requires cultural competency among educators and health professionals, yet knowledge and training in this content is lacking. Thus, we created, piloted, and assessed a six-month interdisciplinary professional development program to enhance knowledge, attitudes, awareness, and cultural humility around LGBTQ+ health for educators, administrators, and clinicians in higher education. Participants were surveyed with standardized assessments pre- and post-program to measure their knowledge, skills, and readiness to teach these topics and incorporate them into their curricula and practice. Participants (n = 33) completed LGBTQ+ inclusivity training including six structured learning sessions, small group mentoring meetings, and a virtual visit to an LGBTQ+ community center. Participants consistently displayed interest and engaged actively in training sessions. In a follow-up survey seven months post-intervention, respondents reported changes in their practice to create inclusive environments. Data analysis demonstrated improved knowledge and comfort with LGBTQ+ care and readiness and willingness to integrate LGBTQ+ care into their curricula and interactions with students. These findings suggest that this interdisciplinary professional development pilot offers a useful program to enable educators and clinicians to support the unique needs of LGBTQ+ communities.