密度梯度离心法和红细胞裂解法对小鼠外周血淋巴细胞表面分子表达的影响  

Effect of Density Gradient Centrifugation and Red Blood Cell Lysis Method on the Expression of Peripheral Blood Lymphocytes Surface Molecules in Rats

作　　者：周扬[1] 钟锐华[1,2] 唐利波[1] 

机构地区：[1]南方医科大学南方医院感染内科,广东广州510515 [2]中山大学附属第五医院感染内科,广东珠海519000

出　　处：《中外医疗》2017年第14期31-33,共3页China & Foreign Medical Treatment

基　　金：广东省自然科学基金(2015A030310440)

摘　　要：目的比较密度梯度离心法和全血红细胞裂解法对小鼠外周血淋巴细胞表面分子表达的影响。方法采用流式细胞术检测密度梯度离心法和全血红细胞裂解法获得的小鼠外周血存活淋巴细胞比例以及表型分子(CD3、CD4、CD8、CD19和CXCR5)的频数。结果密度梯度离心法获得的小鼠外周血存活淋巴细胞比例较全血红细胞裂解法获得的存活淋巴细胞显著升高(92.70%vs 85.90%,P<0.05)。与密度梯度离心法相比,全血红细胞裂解法可使外周血淋巴细胞中CD3、CD4和CD8表达显著下降(60.10%vs 35.90%,P<0.05;42.90%vs 20.30%,P<0.05;24.60%vs 14.80%,P<0.05),而CD19和CXCR5表达则显著升高(12.80%vs 35.30%,P<0.05;15.00%vs 35.10%,P<0.05)。结论密度梯度离心法获得的小鼠外周血淋巴细胞活力强且表型稳定,可作为分离此类细胞的优选方法。Objective To compare the effect of density gradient centrifugation and red blood cell lysis method on the expression of peripheral blood lymphocytes surface molecules in rats.Methods The frequency of survival lymphocyte ratio and surface molecules（CD3,CD4,CD8,CD19 and CXCR5） in rats were tested by the density gradient centrifugation and red blood cell lysis method.Results The peripheral blood survival lymphocytes ratio tested by the density gradient centrifugation was obviously increased compared with that of red blood cell lysis method（92.70% vs 85.90%,P〈0.05）,and the red blood cell lysis method could make the CD3,CD4 and CD8 in the peripheral blood lymphocytes obviously decreased compared with that of density gradient centrifugation（60.10% vs 35.90%,P〈0.05;42.90% vs 20.30%,P〈0.05;24.60% vs14.80%,P〈0.05）,but the expressions of CD19 and CXCR5 obviously increased（12.80% vs 35.30%,P〈0.05;15.00% vs35.10%,P〈0.05）.Conclusion The vigor of peripheral blood lymphocytes of rats obtained by the density gradient centrifugation is strong with steady phenotype,which can be used as the preferred method in isolating this type of cells.

关 键 词：流式细胞术 小鼠外周血 淋巴细胞 密度梯度离心法 全血红细胞裂解法 

分 类 号：R446[医药卫生—诊断学]