高端流式分选与免疫磁珠法纯化T细胞的比较

摘要

Objective To establish an efficient,highly active sorting method for isolating and purifying T lymphocytes;To compare Fluorescence-activated cell sorting (FACS) with Magnetic-activated cell sorting (MACS) in isolating CD8+ T lymphocytes in the spleen of mice.Methods The evaluation was based on the cytologic purity,recovery,activity,morphology and time consumption in the 2 sorting methods.Results (1) The purification rate of CD8+ T lymphocytes was (99.5 ± 0.00) ％ in FACS and (95.6 ± 0.25) ％ in MACS.(2) The recovery rate of CD8+ T lymphocytes was (74.33 ± 3.28) ％ in FACS and (60.00 ± 1.53) ％ in MACS.(3) The time consumption was 1.5 h in FACS and 2.5 h in MACS.(4) The activity ofT lymphocytes was higher in FACS than in MACS.(5) The isolated and purified T lymphocytes by FACS emerged colony growth at 12 h and optimum growth at 24 h,while the cells by MACS grew slower than that by FACS.Conclusion The efficiency and cytologic activity of FACS are higher than those of MACS in isolating T lymphocytes.%目的 为建立一种高效、高活性的T细胞的分离纯化方法,分别采用流式细胞分选技术(FACS)及免疫磁珠分选技术(MACS)从小鼠脾脏组织分离纯化CD8+T细胞.方法 通过测定分选后细胞的纯度、回收率、细胞活力、实验耗时及细胞学形态等指标,综合全面地比较了上述两种纯化方法分离CD8+T细胞的差异.结果 流式细胞分选仪分选后CD8+T细胞纯度为(99.5±0.00)％,而免疫磁珠法的纯度为(95.6±0.25)％.流式细胞分选仪分选CD8+T细胞的回收率为(74.33±3.28)％,而免疫磁珠法的回收率为(60.00±1.53)％.流式细胞分选法耗时1.5 h,免疫磁珠法则需要2.5 h.流式细胞分选后的细胞活力要比免疫磁珠法收集的要高.经流式分选后得到的细胞在12h时就开始出现集落生长,24 h时生长最佳,而经免疫磁珠分选后的细胞总体生长趋势较流式分选后的细胞缓慢.结论 流式细胞分选法较免疫磁珠法效率更高,能得到更高活性的细胞.