Abstract: Osteosarcoma is one of the common primary malignant bone tumors and mainly occurs in children and adolescents. Although comprehensive treatments such as surgery combined with chemotherapy have achieved certain progress in local control, the prognosis of metastatic and recurrent osteosarcoma remains unsatisfactory. In recent years, cancer stem cells(CSCs) have been considered to play a key role in the occurrence, progression, drug resistance, and metastasis of osteosarcoma, and have become a research hotspot. This article systematically reviews the biological characteristics, key molecular markers, signal pathway regulatory mechanisms of CSCs in osteosarcoma and their interaction with the tumor microenvironment, and explores their translational value as potential targets for clinical treatment, aiming to provide a theoretical basis and research direction for the precise treatment of osteosarcoma.