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为了研究急性髓细胞白血病 (AML)细胞诱导成树突状细胞 (DC)的方法及其DC的功能 ,分离 2 5例AML患者骨髓或外周血单个核细胞 ,在含有细胞因子rhGM CSF ,rhIL 4 ,rhTNF α的IMDM完全培养基中培养 8- 12天 ,进行细胞形态学、表型、遗传学及功能测定。结果表明 :2 0 2 5例自诱导培养的第 3天起 ,在倒置显微镜下可观察到部分细胞体积增大 ,形态由圆形变得不规则 ,可见驼峰样或细刺状胞浆突起 ;在第 8- 9天 ,该类细胞所占的比例达到峰值。至第 12天 ,细胞总数及上述形态的细胞数量明显减少。诱导培养结束时收集细胞 ,应用流式细胞术检测 11 2 0例诱导前后细胞表面标志的表达 ,结果表明诱导前细胞不表达CD1a、CD80及CD83,低表达CD86、CD5 4和HLA DR ;诱导后细胞CD1a、CD80、CD83、CD86、CD5 4及HLA DR的表达明显上调。在同种异体混合淋巴细胞反应 (Allo MLR)中 ,该类细胞刺激同种异体淋巴细胞增殖—对3H TdR的摄取能力明显高于AML细胞。FISH证实诱导生成的具有DC特征细胞的AML起源。结论 :体外联合应用细胞因子可将AML细胞诱导成具有DC形态、表型及功能特征的细胞 (AML DC)。
In order to study the method of DC induced by acute myeloid leukemia (AML) and its function in DC, 25 bone marrow or peripheral blood mononuclear cells from AML patients were isolated and cultured in DMEM containing rhGM CSF, rhIL 4 , rhTNF α IMDM complete culture medium for 8-12 days, cell morphological, phenotypic, genetic and functional assays. The results showed that after 20 05 self-induced culture on day 3, the volume of some cells was observed under an inverted microscope, and the shape of the cell became irregular from the round shape, showing the appearance of the hump-like or spiculate cytoplasm; On days 8-9, the proportion of such cells peaked. By the 12th day, the total number of cells and the number of cells in the above morphology decreased significantly. The cells were harvested at the end of induction culture, and the cell surface markers were detected by flow cytometry before and after induction. The results showed that the cells did not express CD1a, CD80 and CD83 before induction, but low expressed CD86, CD54 and HLA DR The expression of CD1a, CD80, CD83, CD86, CD5 4 and HLA DR were significantly up-regulated. In allogeneic mixed lymphocyte reaction (Allo MLR), these cells stimulate proliferation of allogeneic lymphocytes - uptake of 3H TdR is significantly higher than AML cells. FISH confirmed the induction of AML-derived DCs with AML origin. CONCLUSION: AML cells can be induced into cells with DC morphology, phenotype and functional characteristics (AML DC) by combining cytokines in vitro.