摘要
目的：比较噻唑兰（MTT）法和CCK-8法检测小鼠淋巴细胞增殖反应的最佳实验条件,并比较两种不同检测方法的灵敏度.方法：分离小鼠脾脏淋巴细胞,分别比较不同培养时间（69、70、71、72、73 h）、不同反应剂量（5、10、15、20、25 μl）、不同细胞数量（0、2.5×10^4、5×10^4、10×10^4、20×10^4）反应体系的MTT法和CCK-8法的吸光度值.结果：对于2×105的反应体系,MTT法的最佳培养时间为71 h,最佳反应剂量为15 μl.而CCK-8法的最佳培养时间为72 h,最佳反应剂量为10 μl.同时,通过对比两种不同检测方法对于相同细胞数量培养体系的吸光度值发现,CCK-8法的灵敏度要高于MTT法.结论：CCK-8法是一种较MTT法更优的检测小鼠淋巴细胞增殖反应的实验方法.
Objective：To compare the optimal experimental condition and different sensitivity in murine lympho-cyte proliferation response as measured by thiazolyl blue （MTT）and cell counting kit-8 （CCK-8）assays. Method： Spleen lymphocytes were isolated from mice, and subsequently lymphocyte proliferative activities were compared with different culture intervals, dosages, and cell counts, respectively. Results： For 2 ×10^5 lymphocytes reaction system, the best incubation time of MTT was 71 hours, and the best reaction dose was 15 μl. For CCK-8 assay, the best incubation time of CCK-8 was 72 hours, and reaction dose was 10 μl. In addition, by comparing the pro-liferative activities measured by the two different methods for the same cell culture system, it was shown that the sensitivity of CCK-8 assay was much higher than that of MTT assay. Conclusion： CCK-8 assay appears to be a better method for measurement of lymphocyte proliferation response in mice.
出处
《感染．炎症．修复》
2012年第4期199-202,共4页
Infection Inflammation Repair
基金
国家自然科学基金资助项目(30971192
81071545)
国家重点基础研究发展计划项目(201 2CB518102)
全军"十二五"计划重大项目资助(AWS11J008)
关键词
噻唑兰
CCK-8
淋巴细胞
增殖反应
Thiazolyl blue
Cell counting kit-8
Lymphocyte
Proliferation response
