首先用Percoll连续密度梯度离心法制备外周血单个核细胞,采用流式细胞仪分选CD3+CD4+T淋巴细胞。分选后的细胞通过细胞存活率、细胞纯度鉴定和形态观察对分选方法进行评价。结果显示,Percoll连续密度梯度离心法能很好的制备外周血单个核细胞,分选前CD3+CD4+T淋巴细胞纯度为(51.9±9.6)%,分选后CD3+CD4+T淋巴细胞纯度为(95.9±0.8)%,差异有统计学意义(P〈0.01)。分选后的细胞存活率为(95.8±1.2)%,细胞的形态保持完整。
First of all, peripheral blood mononuelear cells (PBMC) were prepared by using continuous density Percoll gradient centrifugation technique, and then CD3 + CD4+T lymphocytes were sorted by flow cytometry (FCM). The method was evaluated by the purity, shape and the survival rate of the sorted cells, respectively. The result shows that continuous density Percoll gradient centrifugation could well prepare PBMC. The purity of CD3 + CD4 + T lymphocytes was (51.9 ± 9.6) % before sorting, while the purity of CD3 + CD4 + T lymphocytes was (95.9 ± 0. 8 ) % after sorting, it was significantly different between before and after sorting ( P 〈 0.01 ). The sorted CD3 + CD4 + T lymphocytes survival rate was (95.8 ± 1.2) % and had intact cell shape.