MTT法检测大鼠外周血淋巴细胞增殖反应探讨与应用

发布时间：2018-02-21 06:02

本文关键词： MTT法淋巴细胞增殖细胞免疫　出处：《中国农学通报》2016年26期 　论文类型：期刊论文

【摘要】：为了更精准简捷地评价机体的细胞免疫状态,采用MTT比色分析法检测大鼠外周血淋巴细胞的增殖能力,并对试验的最适条件进行了探讨,然后将该法应用到Transwell共培养体系中以评价H9N2AIV感染内皮细胞后对跨内皮淋巴细胞增殖能力的影响。结果表明:在96孔细胞培养板中接种密度为1.25×106个/m L的淋巴细胞于含10%FBS的RPMI 1640培养基中培养的最适时间为48 h。感染病毒后,跨内皮淋巴细胞的增殖明显受抑制;内皮完整状态下,淋巴细胞和病毒共孵育后,活病毒的量降低,而当内皮不存在或被病毒感染后,这一现象消失。结果证明作为机体细胞免疫功能评价指标之一的淋巴细胞增殖能力可采用改良后的MTT法检测。
[Abstract]:In order to evaluate the cellular immune status more accurately and simply, the proliferative ability of peripheral blood lymphocytes in rats was detected by MTT colorimetric assay, and the optimum conditions of the test were discussed. Then the method was applied to the Transwell co-culture system to evaluate the effect of H9N2AIV infection on the proliferation of transendothelial lymphocytes. The results showed that lymphocytes with a density of 1.25 脳 106 / mL were inoculated in a 96-well cell culture plate. The optimal culture time in RPMI 1640 medium containing 10s was 48 h. The proliferation of transendothelial lymphocytes was obviously inhibited. When the endothelial cells were intact, the amount of live virus decreased after the lymphocytes were incubated with virus, but when the endothelial cells did not exist or were infected by the virus, The results showed that the lymphocyte proliferation ability, as one of the evaluation indexes of cellular immune function, could be detected by improved MTT method.
【作者单位】：北京农学院动物科学技术学院/兽医学(中医药)北京市重点实验室;中国农业大学动物医学院;
【基金】：国家自然科学基金“抗菌中药方剂清除感染细菌的新机制研究”(31272144)
【分类号】：S852.4

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