Background The present study intends to shed light on the (inter)subjective and professional modalities through which prison workers manage to cope with the presence of incarcerated transgender people in Italy. Method Fourteen semi structured interviews were undertaken with prison-workers in three Italian jails. The transcribed interviews were processed through text-mining-analysis, and an in-depth investigation of these discursive sources was performed to access the sociocultural representation of prison-workers concerning transgender related issues. Results Findings showed different discursive positionings, which vary significantly according to the professional role and gender identity of the interviewees. Although specialized training aimed at improving knowledge on transgender issues and interpersonal skills had been previously provided to all prison-workers, non-transgender affirming behaviors and attitudes continue to be observed. However, they may be the product of the persistence of cultural biases belonging to the more general societal context. Conclusions Specific training programs addressed to prison-workers are certainly necessary, but their real effectiveness should be constantly monitored and verified. At the same time, it seems that an effective improvement of the condition of transgender people within correctional facilities may only be obtained through paradigmatic changes concerning representations about gender nonconformity within the more general Italian culture and society.