南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (05): 599-602.

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标记后转输的凋亡细胞流式细胞术体内示踪方法

汪艳； 高毅； 孙尔维； 谢金敏； 张会迎； 陈剑波；   

  1. 南方医科大学珠江医院普外科； 南方医科大学珠江医院肾移植科移植免疫研究所； 南方医科大学中心实验室 广东广州510282； 广东广州510282； 广东广州510515；
  • 出版日期:2006-05-20 发布日期:2006-05-20
  • 基金资助:
    广东省自然科学基金重点项目(037055);国家自然科学基金(30571757)~~

In vivo tracing of transferred apoptotic cell labeled using CFSE:a flow cytometry-based assay method

WANG Yan1,GAO Yi1,SUN Er-wei2,XIE Jin-min1,ZHANG Hui-ying1,CHEN Jian-bo3 1Department of General Surgery,2 Institute of Transplantation Immunology,Department of Kidney Transplantation,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China;3Central Laboratory,Southern Medical University,Guangzhou 510515,China   

  1. 南方医科大学珠江医院普外科； 南方医科大学珠江医院肾移植科移植免疫研究所； 南方医科大学中心实验室 广东广州510282； 广东广州510282； 广东广州510515；
  • Online:2006-05-20 Published:2006-05-20

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摘要： 目的探索一种可用于转输凋亡细胞体内示踪研究的方法。方法首先对分离所得活细胞进行(CFSE)荧光标记,再诱导细胞凋亡。将该细胞经静脉注射入受体后,采用流式细胞术和组织学荧光显微镜观察标记凋亡细胞在不同组织的动态分布。同时进行CFSE标记稳定性观察实验。结果CFSE标记凋亡细胞的阳性率高,荧光强度大,不会迅速衰减和外漏。流式细胞术检测可发现标记凋亡细胞在不同组织的动态分布。结论CFSE标记细胞后诱导凋亡,再经流式细胞术检测组织内细胞分布,必要时辅以组织学检测可以作为一种可靠、高效、经济的凋亡细胞体内示踪研究技术方法。

Abstract: Objective To establish an assay method for detecting the migration of transferred apoptotic cells into the recipient using flow cytometry.Methods Spleen lymphocytes were isolated and labeled with an intracellular amine dye,carboxyfluorescein diacetate succinimidyl ester(CFSE),to allow discrimination.The labeled cells were induced with dexamethasone to undergo apoptosis and transferred into recipient mice via tail venous transfusion.Flow cytometry and histological examination of different tissues were performed at different time points.The stability of CFSE labeling for apoptotic cells was also tested.Results The CFSE-labeled apoptotic cells were highly fluorescent with a positive labeling rate of(98.0±1.9)%.The stability of CFSE-labeling was testified,and the CFSE-labeled apoptotic cells entering different tissues at different time points were detected by flow cytometry and verified by histological examination.Conclusion Flow cytometry using CFSE labeling is reliable,sensitive,precise and convenient for apoptotic cell tracing in vivo and in vitro.

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