MTT法检测大鼠外周血淋巴细胞增殖反应探讨与应用

doi:10.11924/j.issn.1000-6850.casb16060080

中国农学通报 ›› 2016, Vol. 32 ›› Issue (26): 21-26.doi: 10.11924/j.issn.1000-6850.casb16060080

所属专题：生物技术；畜牧兽医

MTT法检测大鼠外周血淋巴细胞增殖反应探讨与应用

梁宏伟¹,朱雯宇¹,冯波^1,2,王建舫¹,张涛¹,穆祥¹

(¹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,北京 102206；²中国农业大学动物医学院,北京 100193）

收稿日期:2016-06-16 修回日期:2016-09-11 接受日期:2016-08-30 出版日期:2016-09-21 发布日期:2016-09-21
通讯作者: 穆祥
基金资助:
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)。

Application of MTT Method for Detecting Rat Peripheral Blood Lymphocyte Proliferation

Liang Hongwei¹, Zhu Wenyu¹, Feng Bo^1,2, Wang Jianfang¹, Zhang Tao¹, Mu Xiang¹

(¹Beijing Key Laboratory of Traditional Chinese Veterinary Medicine, Beijing University of Agriculture, Beijing 102206;²College of Veterinary Medicine, China Agricultural University, Beijing 100193)

Received:2016-06-16 Revised:2016-09-11 Accepted:2016-08-30 Online:2016-09-21 Published:2016-09-21

摘要/Abstract

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H9N2 AIV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/mL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。

关键词: 分子标记辅助选择, 分子标记辅助选择, 优质蛋白玉米, 近等基因系.

Abstract: In order to evaluate cellular immune state more accurately and simply, we used MTT colorimetric analysis to detect the proliferation of peripheral blood lymphocytes in rats, and analyzed the optimum conditions of the experiment. Then we applied MTT method to the Transwell co-culture system in order to assess the effect of infecting endothelial cells by H9N2 AIV on the proliferation of lymphocytes which crossing endothelial cells. Results showed that the optimum culture time was 48 h when inoculating 1.25×10⁶ lymphocytes per milliliter into the RPMI 1640 medium with 10% FBS in a 96 hole cell culture plate. The proliferation of lymphocytes was inhibited apparently after infected by virus. Lymphocytes and virus co-incubation reduced the amount of live virus with intact endothelium. On the contrary, this phenomenon disappeared. The result indicated that as one of the evaluation indexes of cellular immunity, lymphocytes proliferation could be detected by the optimized MTT method.

梁宏伟,朱雯宇,冯波,王建舫,张涛,穆祥. MTT法检测大鼠外周血淋巴细胞增殖反应探讨与应用[J]. 中国农学通报, 2016, 32(26): 21-26.

Liang Hongwei,Zhu Wenyu,Feng Bo,Wang Jianfang,Zhang Tao and Mu Xiang. Application of MTT Method for Detecting Rat Peripheral Blood Lymphocyte Proliferation[J]. Chinese Agricultural Science Bulletin, 2016, 32(26): 21-26.

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MTT法检测大鼠外周血淋巴细胞增殖反应探讨与应用

Application of MTT Method for Detecting Rat Peripheral Blood Lymphocyte Proliferation

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