Purpose: Knowledge of normal anatomy and variations of testicular arteries are critical for planning before minimally invasive surgery. In this article, we aimed to evaluate the morphological characteristics of testicular arteries by MDCT and propose a new classification system. Method: A total of 400 adult patients (total of 819 testicular arteries) who underwent abdominal CT angiography examination for several unrelated reasons were enrolled in this study. Testicular arteries (TAs) were evaluated in terms of number, origin, course, and caliber. Based on our findings, we also proposed a novel anatomical classification for the TAs according to their origin (divided into 5 types-type 1-5-) and course (divided into three types-type a-c-). Type 4 origin and type a course are the expected normal origin and course of bilateral TAs. Results: The most common type based on the origin was type 4, with the testicular artery originating from the abdominal aorta, inferior to the level of the renal artery within 5 cm distal to the renal arteries. 70.7% of the right and 75.6% of the left testicular arteries were type 4. The most common type of the vessel course was the normal expected trajectory (type a) with an incidence of 76.3% for the right and 82.6% for the left testicular arteries. Conclusion: Normal anatomy and variations of testicular arteries may be effectively evaluated by CT angiography in a non-invasive manner.