白蛋白和放置时间对细胞因子诱导杀伤细胞活性的影响及简易检测方法

发布时间：2018-01-06 02:34

本文关键词：白蛋白和放置时间对细胞因子诱导杀伤细胞活性的影响及简易检测方法　出处：《南方医科大学学报》2013年07期 　论文类型：期刊论文

【摘要】：目的研究细胞因子诱导杀伤细胞(CIK)回输液中添加白蛋白和放置时间对其活性的影响,并建立简捷的CD3+CD56+细胞生成率及细胞死亡率的检测方法。方法(1)抗CD3-FITC和抗CD56-PE同时标记CIK细胞,或FQ-AE染色CIK细胞,荧光显微镜观察并记录;(2)悬浮液中加或不加白蛋白,Annexin V-FITC标记CIK细胞,不同时间段取样,流式细胞仪检测。结果(1)食道癌患者CIK细胞集落小,散在分布;胰腺癌患者CIK细胞集落大且集中;(2)显微镜可观察并计数CD3+CD56+细胞;(3)回输液中加与不加白蛋白的凋亡率及死亡率均无显著差异;(4)CIK细胞在培养箱外放置,0～90 min之间样品与150 min样品之间凋亡细胞数差异显著,细胞死亡率与放置时间无显著差异。结论(1)不同肿瘤患者CIK细胞的生长集落不同;(2)抗CD3和抗CD56标记可计数CD3+CD56+CIK细胞生成率,FQ-AE染色可计数死亡细胞比率,检测方法简便、快捷;(3)CIK细胞回输液中可不加白蛋白,对细胞存活率无影响;培养箱外放置时间90 min之内为宜,时间越短越好。
[Abstract]:Objective to study the effects of albumin addition and storage time on the activity of cytokine induced killer cell line CIK. A simple method for the detection of cell growth rate and cell death rate of CD3 CD56 was established. Methods 1) both anti CD3-FITC and anti CD56-PE were used to label CIK cells. Or FQ-AE stained CIK cells were observed and recorded by fluorescence microscope. (2) CIK cells were labeled with or without Annexin V-FITC in suspensions and sampled at different time intervals. Results 1) the colony of CIK cells in patients with esophageal cancer was small and scattered. The colony of CIK cells in pancreatic cancer patients was large and concentrated. (2) CD3 CD56 cells could be observed and counted by microscope. (3) there was no significant difference in apoptosis rate and mortality rate between adding and not adding albumin. There was significant difference in the number of apoptotic cells between the #number0# min sample and the 150 min sample placed outside the incubator. There was no significant difference between cell death rate and storage time. Conclusion (1) the growth and colony of CIK cells in different tumor patients were different. (2) Anti CD3 and anti CD56 labeling can count CD3 CD56 CIK cell formation rate and FQ-AE staining can count the rate of dead cells, the detection method is simple and fast; No albumin was added to the transfusion of CIK cells, which had no effect on the survival rate of CIK cells. The outside of the incubator should be placed within 90 min, the shorter the better.
【作者单位】：广州医学院附属肿瘤医院肿瘤科;
【基金】：广东省医学科研基金(A2011285) 广州医学院基金(编号:28)
【分类号】：R392.1
【正文快照】： 细胞因子诱导的杀伤细胞(CIK)是一群由IFN-γ、IL-2及抗CD3单抗诱导的多克隆异质细胞,可在体内外杀伤多种肿瘤细胞[1-2]。因其有令人鼓舞的疗效和较轻微的毒副作用[3],国内外很多医院已应用于肿瘤临床治疗。最初,CIK是指正常人体外周血中只占1%~5%的CD3+CD56+细胞,目前国内外

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