应用Percoll分离纯化NK细胞  

ISOLATION AND PURIFICATION OF HUMAN NK CELLS BY PERCOLL

作　　者：邬于川[1] 凌体淑[1] 刘玉林[2] 李程旭[1] 赵世鸿[1] 

机构地区：[1]泸州医学院微生物学与免疫学教研室,泸州646000 [2]泸州医学院附属医院麻醉科,泸州646000

出　　处：《泸州医学院学报》2000年第2期102-104,共3页Journal of Luzhou Medical College

基　　金：四川省教委资助

摘　　要：目的 :从人外周血中分离纯化高浓度、高纯度及活性良好的静息状态NK细胞 ,为进一步研究其特性打下基础。方法 :应用Percoll非连续密度梯度离心法结合panning负分离法获得所需NK细胞 ,并用免疫组化Envision二步法、MTT还原法分别对NK细胞的纯度、酶活性及细胞毒活性进行全面鉴定。结果 :NK细胞纯度为 6 0～ 80 % ,浓度最低可达 10 6/ml,细胞酶活性良好 ,细胞毒活性平均达 5 0 %。结论 :此分离法所获NK细胞数量及纯度较高 ,不影响细胞的酶活性 ,也不会激活NK细胞 ,是一种良好的静息状态NK细胞的分离方法。Objective: To separate the resting NK cells with higher yields, higher purity and better viability from human peripheral blood cells. Methods: Human NK cells were acquired by discontinuous percoll gradient centrifugation and panning negative methods and the Envision methods and MTT colormetry were employed to determin the NK cells purity and viability respectively. Results: The purity of acquired NK cells was 60～80%. The concentration could at least be 10 6/ml and the cells had better enzyme viability and killer function. Conclusion: The acquired NK cells had higher yields and purity. Their viability was not influenced by percoll. Therefore, it is believed this separation method is good for the isolation of resting NK cells.

关 键 词：PERCOLL NK细胞 分离 纯化 panning 细胞活性 

分 类 号：R392.12[医药卫生—免疫学]