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目的对急性髓系白血病(AML)骨髓单个核细胞进行体外培养,获得白血病干细胞并进行特征鉴定。方法体外培养AML患者的骨髓单个核细胞,观察其形态学特征、长期存活及增殖特性,用染色体分析和流式细胞分析技术研究其核型和细胞表面标志,用RT-PCR技术分析其nucleostemin、Bmi-1、ABCG2、prominin1、OCT4、Nanog等干细胞相关基因的表达。结果培养的白血病细胞呈集落样悬浮生长,在体外能存活12个月以上并保持增殖特性。染色体核型呈亚四倍体。表达上述干细胞相关基因。细胞表面表达CD38、CD29、CD44和HLA-DR,弱表达CD19和CD71,不表达CD34、CD13、CD2和CD105。结论经培养获得的白血病细胞,具备肿瘤干细胞的一些特征,提示为白血病干细胞。
Objective To culture leukemic stem cells from acute myeloid leukemia (AML) bone marrow mononuclear cells in vitro and characterize them. Methods The bone marrow mononuclear cells of AML patients were cultured in vitro. Morphological characteristics, long-term survival and proliferation of AML patients were observed. The karyotype and cell surface markers were analyzed by chromosome analysis and flow cytometry. The nucleostemin, Bmi-1, ABCG2, prominin1, OCT4, Nanog and other stem cell-related gene expression. Results The cultured leukemia cells grew in colony-like suspension and survived for more than 12 months in vitro and maintained the proliferation characteristics. Karyotype was sub-tetraploid. Expression of the above stem cell related genes. Cell surface expression of CD38, CD29, CD44 and HLA-DR, weak expression of CD19 and CD71, does not express CD34, CD13, CD2 and CD105. Conclusion Leukemic cells obtained by culturing have some characteristics of tumor stem cells, suggesting leukemic stem cells.