目的 探索CCK-8法檢測(cè)小鼠脾淋巴細(xì)胞增殖的最佳條件。方法 采用交叉設(shè)計(jì)或兩因素完全隨機(jī)設(shè)計(jì)探索分裂素刀豆蛋白（ConA）或脂多糖（LPS）刺激的脾細(xì)胞增殖實(shí)驗(yàn)的細(xì)胞制備方法、細(xì)胞濃度、分裂素濃度、刺激時(shí)間、培養(yǎng)基中胎牛血清（FBS）濃度、是否預(yù)培養(yǎng)等條件。結(jié)果 制備方法中以輕壓制備的脾細(xì)胞增殖率高于輕磨；脾細(xì)胞濃度以5×10⁶/mL為最佳；ConA濃度為2、5、10 μg/mL，LPS濃度為10、20、50 μg/mL細(xì)胞增殖率未見明顯差異；刺激時(shí)間在48和72 h間脾細(xì)胞增殖率未見明顯差異；用含10%、15%和20% FBS的培養(yǎng)基培養(yǎng)的脾細(xì)胞增殖率未見明顯差異；制備的脾細(xì)胞當(dāng)天刺激比第二天刺激增殖率更高。結(jié)論 ConA和LPS刺激的Balb/C小鼠脾細(xì)胞增殖實(shí)驗(yàn)的最佳條件為，輕壓制備脾細(xì)胞，脾細(xì)胞濃度為5×10⁶/mL，制備當(dāng)天直接種板，ConA的刺激濃度為2~10 μg/mL，LPS的刺激濃度為10~50 μg/mL。

Objective To study the optimum determination conditions for lymphocytic proliferation by CCK-8 method in mice.Methods To study the different influence factors of spleen cell proliferation experiment stimulated by mitogen concanavalin A (ConA) or lipopolysaccharide (LPS), including cell preparation method, lymphocytic density, FBS and stimulating agent concentration in culture medium, and stimulating immediately or 24 h after preparing cell, with cross design or two factor completely randomized design.Results Spleen lymphocytic proliferation rate of preparation method by light suppression was higher than that of the light grind. The appropriate concentration of spleen cells was 5 × 106/mL. The proliferation rate has no significant difference after being stimulated for 48 or 72 h by ConA (2, 5, or 10 μg/mL) or LPS (10, 20, or 50 μg/mL) under 10%, 15%, or 20% FBS concentration in culture medium. The proliferation rate of stimulating immediately after preparing cell was higher than that of 24 h after preparing cell.Conclusion The optimum conditions of Balb/C mouse spleen cell proliferation assay stimulated by ConA and LPS are as follows:preparation of spleen cells with light pressure, spleen cell concentration of 5×10⁶/mL, direct stimulation with 2-10 μg/mL ConA or 10-50 μg/mL LPS in the day of preparation.