MTT法检测卵蛋白刺激哮喘小鼠脾淋巴细胞增殖最佳实验条件的研究

发布时间：2018-02-15 19:31

本文关键词： MTT法哮喘脾淋巴细胞卵蛋白　出处：《毒理学杂志》2014年03期 　论文类型：期刊论文

【摘要】：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(r0.99)。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。
[Abstract]:Objective to investigate the optimal experimental conditions for the detection of ovalbumin (OVA) in asthmatic mice by MTT. Methods the model of asthmatic mice induced by OVA was established and splenic lymphocytes were isolated and inoculated with different concentrations of splenic lymphocytes. The absorbance A value was measured at different culture time points. The optimal cell concentration and detection time were determined. The effects of different concentrations of OVA on the proliferation of splenic lymphocytes were observed. Results the optimal concentration of splenic lymphocyte inoculation was 2 脳 10 6 + 4 脳 10 6 cells / ml, the best detection time was 48 h. There was a linear relationship between the cell concentration of 1 脳 106 cells / ml and the cell concentration. R 0.99 渭 g / ml OVA could induce the proliferation of splenic lymphocytes significantly, 100 渭 g / ml OVA could inhibit the proliferation of splenic lymphocytes. Conclusion the method of MTT can be used to detect the proliferation of splenic lymphocytes in asthmatic mice. There was a linear relationship between cell concentration and A value. The concentration of OVA was an important factor affecting the proliferation of splenic lymphocytes.
【作者单位】：第三军医大学新桥医院全军呼吸疾病研究所;
【基金】：国家自然科学基金(30900657) 第三军医大学科研创新基金(2008D203)
【分类号】：R562.25

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