Professionalism stands as a fundamental cornerstone within the realm of physician training, representing a core competency that holds significant importance. It entails creating workplaces that are physically and psychologically safe within the health care system. Positive role modeling from health professionals is important in creating a learning environment that fosters diversity, equity, and inclusion for all. Using the mixed-methods sequential design, this study investigated unprofessional behaviors and biased perceptions demonstrated by health professionals as perceived and experienced by medical students during their clinical rotations at one medical school. Seventy-three M3 students and 37 M4 students responded to the survey, followed by three focus groups (n = 11) to further examine unprofessionalism and biases as well as impacts on performance, learning opportunities, and well-being. The results from both the quantitative and qualitative data demonstrate the presence of unprofessionalism and biases within the current medical education environment. These issues include a lack of respect and compassion, a lack of commitment to professional duties, malfunctioning teamwork, and a lack of sensitivity towards individuals regardless of their group affiliations. The unprofessional behaviors and biased perceptions have detrimental impacts on students’ performance, learning, and well-being. The reasons behind unprofessionalism and bias are multifaceted, influenced by societal and local environmental factors that extend beyond individual beliefs and values. By collecting empirical data on students’ experiences and perceptions, the study sheds light on the areas that need improvement and offers insights into adopting strategies to decrease unprofessional conducts and foster a respectful and inclusive learning environment.