CCK-8法在淋巴细胞增殖检测中最佳实验条件的筛选

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Abstract Objective To optimize the experimental conditions of CCK-8 in lymphocyte proliferation assays. Methods An orthogonal test was designed to investigate the influence of four major factors (cell density, culture period, concentration of LPS and duration of incubation with CCK-8) on cell proliferation of human PBMC and mouse splenocyte. ANOVA was carried out to analyze the stimulation indices of all experimental condition combinations. Results The optimal conditions for CCK-8 was as follows: for PBMC, cell density was 2.5×106/mL, culture period was 48 h, concentration of LPS was 1 μg/mL, and duration of incubation with CCK-8 was 4.5 h; and for splenocyte, cell density was 5.0×106/mL, culture period was 48 h, concentration of LPS was 1 μg/mL, and duration of incubation with CCK-8 was 4.5 h. Conclusion The optimized CCK-8 protocol is a sensitive, convenient and stable quantitative method to evaluate lymphocyte proliferation. This result can provide evidence in screening of immunomodulating drugs and investigation of their immunopharmacology.

Key words： CCK-8 PBMC Lymphocyte proliferation Orthogonal test

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	WANG Jin MA Xiaorong▲ ZHANG Wanggang

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WANG Jin MA Xiaorong▲ ZHANG Wanggang. Optimization of the experimental conditions of CCK-8 in lymphocyte proliferation assays[J]. 中国医药导报, 2018, 15(23): 13-16.

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http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2018/V15/I23/13

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