非连续密度梯度离心法分离红细胞的影响因素研究

发布时间：2018-03-15 20:39

  本文选题：非连续密度梯度离心　切入点：梯度分离　出处：《中国实验血液学杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的:研究非连续密度梯度分离法分离红细胞的影响因素,初步建立Percoll分离液在红细胞分离中的最佳操作条件。方法:检测Percoll与1.5 mol/L NaCl比例为9∶1、10∶1、11∶1、12∶1、13∶1渗透压情况,选取等渗比例作为最佳母液(SIP);以不同密度细胞被充分离心沉降分离,较少存在红细胞滞留、拖带现象作为分离效果评判标准,比较不同红细胞悬液浓度、不同离心力、不同离心加减速度、不同温度和不同离心时间的分离效果。结果:50%以下浓度红细胞悬液分离效果优于50%以上浓度红细胞悬液,4 000×g优于3 000和2 000×g的离心效果,2-8的离心加减速度可以获得满意的离心效果,4℃分层效果比室温稳定,20 min优于15和10 min的分离效果。结论:Percoll与1.5mol/L NaCl为12∶1和11∶1的母液可作为等渗母液,优先4℃离心条件离心,加减速度2-8,其离心效果会随着红细胞悬液浓度的降低、离心时间的延长、离心力的增加而获得更好的分离效果
[Abstract]:Objective: to study the factors affecting the separation of red blood cells by discontinuous density gradient separation, and to establish the optimum operating conditions of Percoll separation solution in erythrocyte separation. Methods: the osmotic pressure of Percoll to 1.5 mol/L NaCl was 9: 1: 10: 1: 11: 11: 12: 1: 1: 1. The isosmotic ratio was chosen as the best mother liquid, the cells of different density were separated by full centrifugation and sedimentation, there was less red blood cell retention, the phenomenon of towing was taken as the criterion for evaluating the separation effect, and the different concentration of red blood cell suspension and different centrifugal force were compared. Different centrifugal speed, Results the separation effect of red blood cell suspensions below 50% was better than that of red blood cell suspensions of more than 50% concentration (4 000 脳 g) than that of 3 000 脳 g and 2 000 脳 g / 2 000 脳 g centrifuge suspensions. The results show that the separation effect at 4 鈩，

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