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目的:从人骨肉瘤株U2-OS中分离并鉴定类肿瘤干细胞。方法:用无血清培养法从骨肉瘤细胞株中分离出肿瘤细胞球,流式细胞术检测骨肉瘤细胞株中Stro-1的阳性表达率,MTT法检测其增殖能力,免疫磁珠分选Stro-1阳性细胞,反转录-聚合酶链反应(RT-PCR)法研究Stro-1阳性细胞中Stro-1 mRNA的表达,并将Stro-1阳性细胞和阴性细胞分别接种于NOD/SCID小鼠。结果:U2-OS中CD44的阳性百分率为5.67%,Stro-1阳性细胞形成的细胞球具有增殖和分化的能力,Stro-1阳性细胞中呈现Stro-1 mRNA的高表达,Stro-1阳性细胞能形成骨肉瘤。结论:骨肉瘤细胞株中存在类骨肉瘤干细胞,并具有自我更新和多向分化的能力。
Objective: To isolate and identify tumor-like stem cells from human osteosarcoma strain U2-OS. METHODS: Tumor cell spheres were isolated from osteosarcoma cell lines by serum-free culture. The positive rate of Stro-1 in osteosarcoma cell lines was detected by flow cytometry. The proliferative capacity was measured by MTT assay. -1 positive cells, reverse transcription-polymerase chain reaction (RT-PCR) method to study the expression of Stro-1 mRNA in Stro-1 positive cells, and inoculate Stro-1 positive cells and negative cells in NOD/SCID mouse. RESULTS: The positive percentage of CD44 in U2-OS was 5.67%. Stro-1 positive cells had the ability to proliferate and differentiate, Stro-1 positive cells showed high expression of Stro-1 mRNA, and Stro-1 positive cells. Can form osteosarcoma. CONCLUSION: Osteoid sarcoma stem cells exist in osteosarcoma cell lines and have the ability to self-renew and multi-differentiate.