目的探讨MTT法检测卵蛋白(OVA)刺激哮喘小鼠脾淋巴细胞增殖的最佳实验条件。方法制备OVA诱导哮喘小鼠模型,分离脾淋巴细胞。接种不同浓度的脾淋巴细胞,在不同培养时间点测定吸光度(A)值,确定最佳细胞浓度和检测时间。观察不同浓度OVA对脾淋巴细胞增殖的影响。结果最佳脾淋巴细胞接种浓度为2×106~4×106cells/ml,最佳检测时间为48 h。培养24和48 h时,细胞浓度为1×106~4×106cells/ml,A值与细胞浓度呈线性关系(r>0.99)。10μg/ml OVA可诱导脾淋巴细胞显著增殖,100μg/ml OVA反而抑制脾淋巴细胞增殖。结论 MTT法可用于检测哮喘小鼠的脾淋巴细胞增殖,细胞浓度与A值呈线性关系。OVA浓度是影响脾淋巴细胞增殖的重要因素。 Objective To investigate the best experimental conditions for measuring the proliferation of splenic lymphocytes induced by ovalbumin (OVA) in asthmatic mice by MTT assay. Methods OVA induced asthma mouse model, isolated spleen lymphocytes. Inoculation of different concentrations of splenic lymphocytes, absorbance (A) values ​​measured at different culture time points, to determine the optimal cell concentration and detection time. To observe the effect of different concentrations of OVA on the proliferation of splenic lymphocytes. Results The best splenic lymphocyte inoculum concentration was 2 × 106 ~ 4 × 106cells / ml, the best detection time was 48 h. The cell concentration was 1 × 106 ~ 4 × 106cells / ml at 24h and 48h, and the value of A was linear with cell concentration (r> 0.99). 10μg / ml OVA can induce significant proliferation of splenic lymphocytes, 100μg / ml OVA but inhibit spleen lymphocyte proliferation. Conclusion MTT method can be used to detect the proliferation of splenic lymphocytes in asthmatic mice. The concentration of cells is linear with the value of A. OVA concentration is an important factor affecting the proliferation of spleen lymphocytes.