Objectives. The aim of this qualitative study was to explore the attitudes, beliefs, and preferences of Italian osteopaths regarding the management of transgender patients through a content analysis of emergent data from semi-structured interviews. Methods. This study was a content analysis based on the Standards for Reporting Qualitative Research guidelines. Purposive sampling of 10 Italian osteopaths was applied. Data were collected through semi-structured interviews, from March to April 2021, and subsequently transcribed verbatim with the content analysis carried out as an iterative process. Results. One participant was excluded during the first interview due to them being unsuitable for this study. Data saturation was reached after two interviews with the remaining nine participants. Data analysis revealed four main themes: microaggressions, acceptance and non-judgement, person-centered treatment, and education implementation. Conclusions. This study presents cisgender Italian osteopaths’ attitudes in the care of transgender people, revealing the desire to embrace and apply osteopathic tenets regardless of the patient’s gender identity.