Organoids, an emerging bioengineering approach, can replicate the structure and function of human organs in vitro, providing valuable tools for biomedical research. Patient-derived organoids (PDOs) retain the histological and genetic characteristics of the original tumor, playing a crucial role in precision oncology. Ultrasound-assisted technology, particularly multimodal ultrasound imaging, facilitates the establishment of PDOs for inoperable or rare tumors by enabling precise tumor localization, minimally invasive tissue sampling, and high-purity cell extraction.This review explores the application of multimodal ultrasound techniques in PDO development across various tumor types, highlighting their advantages in obtaining high-purity tumor cells, improving PDO culture success rates, and advancing personalized cancer treatment. Additionally, we discuss factors influencing PDO establishment, such as tumor type, sampling methods, and bioengineering advancements. By summarizing current progress and challenges, this review provides insights into optimizing ultrasound-assisted PDO construction for clinical and research applications.