Objective: By primary cultivation of clinical specimens of human cervical cancer, the main and side population cells were isolated and their biological characteristics were analyzed. Methods: Clinical specimens of human cervical tissue were cultured to get the primary cells. After staining by Hoechst33342, flow cytometry was used for sorting to get the main and side population cells. Their molecular biology, morphology and tumorigenicity were compared with that of Hela cells. Results: The primary cervical cancer and side population cells were isolated successfully, and the two cells have obvious tumor cell morphology. CK17, CK19, P63 and beta 1-integrin and other related markers were expressed. Tumorigenicity of side population cells was significantly higher than the main group of cells. Distribution of the cytoskeleton and expression associated tumor markers were not affected before and after tumorigenesis of the two cells. Conclusion: Side population cells which were cultured and isolated from clinical specimens of human cervical cancer have the characteristics of tumor stem cell, which can be used for further study of targeted therapy for cervical cancer.