免疫磁珠法及密度梯度离心法分离纯化人外周血中性粒细胞的比较

Application of immunomagnetic beads and density gradient centrifugation in isolation of neutrophilic granulocytes from human peripheral blood

目的比较免疫磁珠法和密度梯度离心法分离纯化人外周血中性粒细胞的可靠性。方法用免疫磁珠法和密度梯度离心法分别分离中性粒细胞,使用血球计数仪结合显微镜鉴定细胞纯度、回收率,以台盼蓝检测细胞活力。结果免疫磁珠法与密度梯度离心法分离中性粒细胞纯度分别为(98.76±0.53)%,(92.34±1.57)%,回收率分别为(90.01±2.24)%和(82.20±2.17)%,两种方法分离后中性粒细胞活力分别为(95.60±2.30)%,(94.20±3.11)%。结论免疫磁珠法分离人中性粒细胞纯度及回收率均好于密度梯度离心法,但分离后细胞活性二者差异不大。

Objective To compare the reliability of immunomagnetic beads and density gradient centrifugation in purification and isolation of neutrophilic granulocytes from human peripheral blood. Methods Neutrophilic granulocytes were isolated with immunomagnetic beads and density gradient centrifugation, respectively. Their purity and recovery rate were determined with a blood cell counter in combination with microscopy and cell viability was detected with trypan-blue. Results The purity of neutrophilic granulocytes, isolated with immunomagnetic beads and density gradient centrifugation, was 98.76%± 0.53% and 92.34% ± 1.57% respectively, with a recovery rate of 90.01% ±2.24% and 82.20% ± 2.17% respectively, and a cell viability of 95.60% ± 2.30% and 94.20% ± 3.11% respectively. Conclusion The purity and recovery rate of neutrophilic granulocytes isolated with density gradient centrifugation are better than those of neutrophilic granulocytes isolated with immunomagnetic beads. However, there is no significant difference in cell viability of neutrophilic granulocytes after they are isolated.