The spinal cord is integral to various functions within the central nervous system, including sensory, motor, visceral, and reflex mechanisms. While previous research has identified several neurochemical markers in the spinal cords of rodents, cats, mice, and humans, there is limited information regarding the distribution of the cocaine- and amphetamine-regulated transcript (CART) peptide in the human spinal cord. Additionally, the precise functions of CART in relation to sensory, motor, and limbic system processes remain incompletely understood. Therefore, this study aimed to elucidate the localization of CART in the human spinal cord. Segments of the human C1-Co1 spinal cord were fixed with 10% formalin, then transversely sectioned using a cryostat and stained for CART immunohistochemistry to identify CART localization. CART-immunoreactive (CART-IR) fibers were consistently observed in the anterior, lateral, and dorsal funiculus of the white matter across all spinal cord segments. Within the dorsal horn, dense CART-IR fibers and a small number of neurons were present in laminae I and II, while laminae III and IV exhibited fewer CART-IR fibers and neurons. Moderate numbers of CART-IR fibers and neurons were observed in laminae V, VII, and VIII. Notably, dense CART-IR was detected in neurons of the intermediolateral nucleus in thoracic and upper lumbar spinal cord segments. Laminae IX and X displayed numerous CART-IR neurons and fibers. This study represents the first identification of CART peptide localization within the human spinal cord, shedding light on its distribution and potential functional roles.