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人外周血树突状细胞的体外扩增及鉴定 被引量:91

Generation of Dendritic Cells from Human Peripheral Blood
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摘要 树突状细胞(DC)作为抗原提呈细胞,在激发T细胞免疫应答及T细胞依赖性抗体生成中起重要作用,骨髓及外周血的CD34造血前体细胞在GM-CSF和TNF-α的作用下,可在体外分化发育,生成树突状细胞.本研究从正常人外周血分离获得单核细胞,加入100ng/ml hGM-CSF、500U/ml hIL-4,体外培养1周后,获得大量高纯度的树突状细胞,其高表达MHC-Ⅰ、MHC-Ⅱ类分子及共刺激分子B7-1和CD40,能强烈激发同种异体T淋巴细胞增殖,培养条件以应用自体血清或胎牛血清为最佳;单独应用hGM-CSF只能生成巨噬细胞;在培养末期加入TNFα可促进DC进一步成熟.本研究为DC的深入研究与临床应用奠定了基础. Dendritic cells (DC) are antigen presenting cells (APC) that play critical roles in the initiation of T cell response and development of T cell-dependent antibodies in vivo. CD34_+ hematopoietic progenitor cells of bone marrow and peripheral blood can differentiate into DC when cultured with GM-CSF and TNF-α in vitro. In the present study, we cultured monocytes isolated from human peripheral blood with 100ng/ml hGM-CSF and 500U/ml hIL-4 for one week, and then found that a large number of DC with high purity were gengerated. DC expressed MHC I , MHC II and costimuladng moleculers highly on cell surface and cound stimulate proliferation of allogeneic T lymphocytes. Self serum or fetal calf serum are best for generation of DC. When hGM-CSF was used alone, the monocytes differentiated into macrophages but not to DC. TNF-α could induce further maturation of DC when added in late period of the culture. Generation of DC from human peripheral blood may facilitate further studies on DC and their clinical applications.
出处 《中国肿瘤生物治疗杂志》 CAS CSCD 1997年第4期302-306,共5页 Chinese Journal of Cancer Biotherapy
基金 国家863基金(Z20-01-03)资助
关键词 树突状细胞 单核细胞 体外扩增 鉴定 外周血 dendritic cell monocyte granulocyte/macrophage colony-stimulating factor interleukin-4 MHC costimulating molecules
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