不同样品处理方法对T淋巴细胞亚群及功能分析的影响

Effect of methods for sample treatment on subsets and function analysis of T lymphocytes

目的研究Ficoll-Hypaque密度梯度离心法、先裂解红细胞后标记抗体、先标记抗体后裂解红细胞3种不同处理方法对T淋巴细胞(T细胞)亚群及功能分析的影响。方法采用3种方法制备样本后,采用流式细胞术检测T细胞表面分子及亚群比例,台盼蓝染色检测细胞存活率,Con A刺激检测细胞活性。结果 Ficoll-Hypaque密度梯度离心法与两种裂解红细胞法相比,T细胞亚群比例差异无统计学意义(P>0.05),细胞存活率及细胞活性均较高(P<0.05)。两种裂解红细胞法处理样本结果无差异。结论 Ficoll-Hypaque密度梯度离心法适合研究工作,先标记抗体后裂解红细胞的方法更适合临床检验。

Abstract： Objective To investigate the effect of three methods for sample treatment, i. e. Ficoll-Hypaque discontinuous density gradients as well as erythrocyte lysis before and after antibody labeling, on the subsets and function analysis of T lymphocytes. Methods The samples were prepared by three methods and determined for surface molecules and subsets by flow cytometry, for cell survival rate by trypan blue staining, and for cell activity by Con A stimulation. Results The percentages of subsets of T lymphoeytes in samples treated by erythroeyte lysis showed no significant difference with that by Ficoll-Hypaque discontinuous density gradient centrifugation （P 〉 0. 05）, while cell survival rate and activity were higher （P 〈 0. 05）. However, the test results of samples treated by erythrocyte lysis before and after antibody labeling showed no significant difference. Conclusion Discontinuous density gradient centrifugation was more suitable for scien- tific research, while erythrocyte lysis after antibody labeling for clinical laboratory.