非连续密度梯度离心法分离半滑舌鳎外周血白细胞  

Affected factors on separating peripheral blood leukocytes from Cynoglossus semilaevis by discontinuous density gradient centrifugation

作　　者：郝爽[1,2] 罗璋 张振国[1] 刘肖莲[1] 刘国山[1] 冯守明[1,2] 

机构地区：[1]天津市水产研究所,天津300221 [2]天津农学院水产学院,天津300384

出　　处：《河北渔业》2018年第3期1-4,24,共5页

基　　金：天津市水产局青年科技创新项目(J2016-05)

摘　　要：为研究非连续密度梯度离心法分离半滑舌鳎(Cynoglossus semilaevis Günther)外周血白细胞的影响因素,探索Percoll分离半滑舌鳎外周血白细胞的最佳方法,试验设置了密度梯度为1.015/1.065g/mL、1.020/1.070g/mL、1.025/1.075g/mL、1.030/1.080g/mL、1.035/1.085g/mL的五种分离液,分别于600×g、800×g和1 000×g离心力下进行半滑舌鳎白细胞分离,以所得白细胞密度、白细胞纯度、活细胞率作为评判指标,并对分离的白细胞进行了显微观察、鉴定,比较不同密度梯度分离液、不同离心力的分离效果。结果显示:以1.020/1.070g/mL密度梯度分离液、800×g离心力条件下分离效果最佳,每毫升外周血可分离到白细胞数量平均为(2.485±0.182)×107个,白细胞纯度平均为(97.926±0.679)%,活细胞率平均为(97.037±0.925)%。Based on the studies of the affective factors on the separation of peripheral blood leukocytes(PBL)fromCynoglossus semilaevis by discontinuous density gradient method,and the optimal operating conditions of percoll for the separation of PBL was established.The effects of different centrifugal force(1.015/1.065 g/mL、1.020/1.070 g/mL、1.025/1.075 g/mL、1.030/1.080 g/mL、1.035/1.085 g/mL),separation medium density gradient(600×g、800×g、1 000×g)on PBL separation were examined while the separation method was evaluated according to the density and purity of cells,cell survival rate and other facts.The results showed that PBL could be separated effectively with the number of2.485×107 cell/mL,the purity of 97.926%and the survival rate of 97.037%as the proportion of separation liquid is 1.020/1.070 g/mL and the centrifugal force is 800 g.

关 键 词：非连续密度梯度离心 外周血白细胞 半滑舌鳎(Cynoglossus SEMILAEVIS Günther) 

分 类 号：S917.4[农业科学—水产科学]