BACKGROUND:Effective sorting of prostate cancer stem cels is the basis of experimental studies in prostate cancer developing. The most common sorting method is magnetic-activated cel sorting. OBJECTIVE:To separate CD133+/CD44+ cels in prostate cancer tissues using self-designed magnetic beads folowed by culture, passage and immunological identification. METHODS:Self-designed magnetic microspheres were applied to establish immunomagnetic beads to sort CD133+/CD44+ cels in prostate cancer tissues. The sorted cels were cultured in serum-free medium. The sphere formation, cel morphology, and proliferation ability after cel passage were statisticaly compared between the sorted cels and the normal tumor cel lines. Immunofluorescence detection was performed to detect the expression of specific antibodies. RESULTS AND CONCLUSION:Self-designed immunomagnetic beads had smal diameter and a high-sorting effect. The sorted cels possessed a high capacity of microsphere formation. After cel culture and passage, the cels highly expressed CD133 and CD44 antigens. The sorted cels with no induction had varying shapes and grew vigorously. After induction with transforming growth factor-β, the cultured cels were noted to have a single shape and grow slowly. The cel proliferation ability of sorted cels in these two groups differed significantly from that of the normal cancer cel lines (bothP < 0.05). In conclusion, the CD133+/CD44+ cels sorted from prostate tumor cels possessed cel morphology and function characteristics of stem cels which can provide a basis for extraction and culture of prostate cancer stem cels. Cite this article:Gong R, Li SY, Huo ZX, Ding H, Sun EL. Extraction of prostate cancer stem cels using self-designed magnetic beads. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):36-41.