标记后转输的凋亡细胞流式细胞术体内示踪方法

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．

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

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.