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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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外周血pbmc来源的树突状细胞的快速无血清培养方法及细胞信号转导机制
, PP. 1263-1266
Keywords: 树突状细胞,肿瘤坏死因子-α,钙离子载体,细胞信号转导
Abstract:
目的探讨体外无血清条件下诱导人外周血单核细胞(pbmc)迅速生成树突状细胞(dc)的方法,并初步探讨钙离子载体(ci)诱导分化的信号转导途径与肿瘤坏死因子(tnf)-α所诱导的是否相同。方法分离健康献血者的pbmc,给予无血清培养基及50ng/ml的rhgm-csf过夜培养后,再分别给予100ng/ml的a23187或50ng/ml的tnf-α,或预先加入0.5μg/ml的环胞菌素a(csa)30min后,再加入a23187、tnf-α,共培养40h。于相差显微镜下观察细胞形态的变化,流式细胞仪检测细胞的表面标志,mtt比色法检测不同方法处理的pbmc刺激同种异体t细胞的增殖作用。结果健康献血者的pbmc在无血清条件下,给予rhgm-csf及ci或tnf-α培养40h,均可获得dc的典型形态,包括cd14表达下调、cd83及共刺激分子(cd80、cd86)表达上调,以及较强刺激同种异体t细胞增殖的作用;上述由ci诱导的细胞形态的改变、表面分子的表达以及刺激t细胞增殖的作用均可被csa所抑制。而tnf-α所诱导的细胞形态的改变、表面分子的表达以及刺激t细胞增殖的作用均不受csa影响。结论健康献血者的pbmcs在体外无血清条件下,可以被rhgm-csf及ci或tnf-α迅速诱导成dc,但ci与tnf-α诱导pbmc分化为dc的细胞信号转导途径不同。
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