PURPOSE: To evaluate the safety and efficacy of ultrasound-guided percutaneous core needle biopsy of splenic lesions. MATERIALS AND METHODS: This retrospective study included 30 patients who underwent percutaneous core needle biopsy of their splenic lesions using 18- or 20-gauge needles between January 2001 and July 2016 in a single tertiary care center. The characteristics of the splenic lesions were determined by reviewing the ultrasound and computed tomography examinations. Acquisition rate and diagnostic accuracy were calculated, using pathologic results of the splenectomy specimen, clinical course and/or imaging follow-up as a reference standard. Post-procedure complications were identified from electronic medical records, laboratory findings and computed tomography images. Seventy-three specimens were obtained from the 30 patients and splenectomy was performed in 2 patients. RESULTS: Twenty-nine of the 30 patients had focal splenic lesions, while the remaining patient had homogeneous splenomegaly. Acquisition rate and diagnostic accuracy were 80.0% (24/30) and 76.7% (23/30), respectively. Perisplenic hemorrhage without hemodynamic instability developed in one patient. CONCLUSION: Ultrasound-guided percutaneous core needle biopsy of splenic lesions is a safe method for achieving a histopathologic diagnosis and can be considered as an alternative to splenectomy in patients with a high risk of splenectomy-related complications.