[使用羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）标记的凋亡细胞体内追踪：一种基于流式细胞术的检测方法]
[In vivo tracing of transferred apoptotic cell labeled using CFSE: a flow cytometry-based assay method].
作者信息
Wang Yan, Gao Yi, Sun Er-wei, Xie Jin-min, Zhang Hui-ying, Chen Jian-bo
机构信息
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2006 May;26(5):599-602.
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.
目的
建立一种利用流式细胞术检测转移的凋亡细胞向受体迁移的检测方法。
方法
分离脾淋巴细胞，并用细胞内胺染料羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）进行标记，以便区分。用 dexamethasone 诱导标记细胞发生凋亡，并通过尾静脉输血将其转移到受体小鼠体内。在不同时间点对不同组织进行流式细胞术和组织学检查。还测试了 CFSE 对凋亡细胞标记的稳定性。
结果
CFSE 标记的凋亡细胞具有高荧光性，阳性标记率为（98.0±1.9）%。证实了 CFSE 标记的稳定性，通过流式细胞术检测到不同时间点进入不同组织的 CFSE 标记的凋亡细胞，并经组织学检查验证。
结论
使用 CFSE 标记的流式细胞术在体内和体外对凋亡细胞追踪可靠、灵敏、精确且方便。
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