目的: 建立基于人白血病T细胞株Jurkat E6-1的转移因子(TF)生物活性检测方法。方法: 采用细胞毒性和增殖(CCK-8)法探索不同浓度TF对6-巯基嘌呤(6-MP)处理Jurkat E6-1细胞增殖的影响,并对该方法的精密度进行验证。结果: 20 μg·mL-1 TF可使细胞的生存率明显提高,使6-MP的 IC50由(0.46±0.10)μg·mL-1右移至(1.11±0.30)μg·mL-1;当1.0 μg·mL-1 6-MP处理细胞时,TF的浓度和细胞生存率具有良好的线性关系,r>0.95。不同批次间在剂量-反应曲线的每个浓度水平上,RSD均小于20%。TF的半数有效浓度(EC50)为(28.49±9.60)mg·mL-1,可信限率小于20%。结论: TF可以显著提高暴露于6-MP的Jurkat E6-1细胞生存率,该作用呈剂量依赖性,可为TF生物活性测定提供客观、准确的评价手段。
Objective: To develop a method to assess bioactivity of trasfer factor (TF) based on their protective effcets on Jurkat E6-1 cells. Methods: Proliferative effects of different concentrations of TF on 6-mercaptopurine(6-MP) treated Jurkat E6-1 cells detected by CCK-8 assay and the precision was validated. Results: TF exhibited protective activity to change the inhibitory concentration (IC50) value of 6-MP from(0.46±0.10)μg·mL-1 to (1.11±0.30)μg·mL-1 when treated with 20 μg·mL-1 TF. When cells were treated with 6-MP,there is a good linear relationship between the concentration of TF and cell survival rate, r>0.95. At each concentration level of the dose-response curve, the RSD was less than 20%. The median effective concentration (EC50) of TF was (28.49±9.60)μg·mL-1, and the confidence limit was less than 20%. Conclusions: TF significantly improved Jurkat E6-1 cell survival rate in dose dependent manner when challenged with 6-MP. It may be suitable for evaluate bioactivity of TF.
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