The present study describes a case of a spontaneous, unilateral Sertoli cell tumour (SCT) in a 6-year-old pet rabbit. The rabbit was presented with a palpable, unilateral, subcutaneous left inguinal mass, consistent with the suspected clinical diagnosis of neoplasia developing within the retained testis in the inguinal canal. The intrascrotal contralateral testis was palpable, but reduced in volume. The rabbit underwent orchiectomy and both the testes were collected, formalin-fixed, and submitted for histopathological examination. Microscopically, the enlarged testis was effaced by an intratubular SCT in which numerous intratubular microliths were evident. The contralateral testis was severely atrophied. Immunohistochemical stains showed neoplastic Sertoli cells that were diffusely positive for vimentin and anti-Müllerian-Hormone and multifocally positive for cytokeratins and desmin. Eighteen months after the surgery, the rabbit showed no clinical signs of disease. This is the first report of a spontaneously occurring rabbit SCT histologically described and immunohistochemically investigated.