CD4＋T淋巴细胞分选方法的效果评价

目的 建立一种准确且对细胞活性影响小的分离人外周血CD4＋T细胞的分选方法.方法 对人外周血依次采用密度梯度离心（density gradient centrifugation,DGC）和免疫磁珠分选（magnetic-activated cell sorting,MACS）进行CD4＋T细胞分选.分选后的细胞用流式细胞仪（flow cytometry,FCM）、倒置显微镜及细胞周期分析进行分选纯度和生长状态检测.结果 Percoll密度梯度离心法收集的单个核细胞中CD4＋T细胞纯度为（38.8±2.7）％,免疫磁珠分选后CD4＋T细胞纯度为（96.2±0.7）％,两者比较差异有统计学意义（P＜0.01）,且分选后细胞形态及功能完好,24 h内活性最高.结论 密度梯度离心和免疫磁珠分选相结合可以有效提高CD4＋T细胞的分选纯度及保持其活性,可继续用于相关功能研究.

Effect evaluation on sorting method of CD4+T lymphocytes

Objective To establish an accurate sorting method with less interference to the cell activity for isolating CD4＋T lymphocytes.Methods Human peripheral blood cells were sorted by density gradient centrifugation （DGC） and magnetic activated cell sorting （MACS） in turn,and then the cell purity,morphological observation and survival rate of CD4＋T lymphocytes were adopted to conduct the evaluation on the sorting method.Results The purity of CD4＋T lymphocytes was （38.8 ± 2.7）％,while the purity of CD4＋T lymphocytes was （96.2 ± 0.7）％ after magnetic activated cells being sorted,the difference between them was statistically significant （P ＜ 0.01）.The activity was the highest within 24 hours with intact cell shape and function.Conclusion The Percoll DGC combined with the MACS can collect highly pure CD4＋T lymphocytes with less influence on the cell activity and shape,the sorted cells can continue to be used for related researches.