免疫磁珠法在分离纯化外周血CD4^+和CD8^+ T淋巴细胞亚群中的应用

APPLICATION OF MAGNETIC ACTIVARED CELL SORTING FOR SEPARATION AND PURIFICATION OF CD4^+ T CELL AND CD8^+ T CELL SUBPOPULATIONS OF PERIPHERAL BLOOD

目的:探讨免疫磁珠法(MiniMACS)在分离纯化外周血CD4+和CD8+T淋巴细胞亚群中的应用。方法:应用密度梯度离心法分离外周血单个核细胞(Peripheral blood mononucleate cells,PBMC),采用MiniMACS分别分离和纯化39例标本外周血PBMC中的CD4+T淋巴细胞和CD8+T淋巴细胞,并经流式细胞仪分析细胞纯度和台盼蓝染色的方法对细胞活力进行评估。结果:MiniMACS分离外周血CD4+T淋巴细胞前、后细胞纯度分别为(37.38±5.74)%、(97.75±1.03)%(P<0.001),CD8+T淋巴细胞分离纯化前后细胞纯度分别为(20.11±6.83)%、(96.85±1.86)%(P<0.001);外周血分离前PBMC细胞活力为(97.66±2.73)%,纯化为CD4+T淋巴细胞和CD8+T淋巴细胞后细胞活力分别为(97.44±3.08)%、(98.05±2.92)%(P>0.05)。结论:MiniMACS可以高度富集CD4+T淋巴细胞和CD8+T淋巴细胞且不改变细胞的活力。

Objective： To evaluate the separation and purification of T lymphocyte subsets by immunomagnetic beads. Methods： Peripheral blood mononucleate cells （PBMC） were isolated using lymphocytes separation medium and desity gradient centrifugal action. CD4＋T and CD8＋T lymphoeytes were isolated and purified from PBMC by magnetic activated cell sorting （MiniMACS）. The purity and activity of CD4＋T and CD8＋T cells were measured using flow eytometry, trypan-blue dye exclusion test, etc. Results： The results showed that the percentage of CD4＋ T cells before and after the purification was （37.38 ± 5.74） and （97.75 ± 1.03）% （ P 〈0. 001） ; the pereentage of CD8＋T cells before and after the purification was （20.11 ± 6.83） % and （96.85 ± 1.86） % （ P 〈0. 001）, resprectively; and the cell ability was not affected by purification procedure （ P 〈0.05）. Conclusion： MiniMACS can sort out effectively CD4＋T and CD8＋T cell subpopulations of peripheral blood and does not affect the cell vitality.