免疫磁珠及尼龙毛分离纯化T淋巴细胞的比较研究

Comparative research of isolating T lymphocycte population with immunomagnetic beads and nylon wool cells

目的 :探讨免疫磁珠及尼龙毛分离纯化外周血T淋巴细胞的方法及其特点。方法 :用密度梯度离心法分离人外周血单个核细胞 ,再分别以尼龙毛和免疫磁珠分离出T细胞 ,然后使用流式细胞仪鉴定细胞纯度、回收率 ,以台盼蓝检测细胞活力 ,MTT法检测细胞增殖性能。结果 :免疫磁珠方法分离后T细胞的纯度显著高于尼龙毛分离方法 (P <0 .0 5 ) ,而且回收率也大于尼龙毛分离方法 (P <0 .0 0 1 ) ,但二种方法分离的T细胞在细胞活力和增殖性能方面比较差异无显著性 (P >0 .0 5 )。结论 :在细胞生物学研究中 ,免疫磁珠分离法是有效分纯T细胞的较佳方法。

Objective:To evaluate the effect and characteristics of two methods for purifying T lymphocytes.Methods:Human peripheral blood mononuclear cells (PBMC) were prepared with Ficoll-Hypaque density gradient centrifugation, human peripheral T lymphocytes was separated with nylon wool and Dynal immunomagnetic beads isolation,respectively. The purity、viablilty and cell proliferation of T lymphocytes were determined with flow cytometry,trypan blue exclusion and MTT,respectively,then these data were compared with T-test.Results:The purity of T lymphocytes,after isolation with Dynal immunomagnetic beads method(P<0.05) and recovery rate(P<0.001) were significantly higher than that with nylon wool. There was no significant difference in viablilty and cell proliferation activity(P>0.05).Conclusion:Dynal immunomagnetic beads isolation is a very effective and practical method to isolate T cells population from PBMCs in cell biological research.