[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.


CONCLUSIONS
Flow cytometry using CFSE labeling is reliable, sensitive, precise and convenient for apoptotic cell tracing in vivo and in vitro.
Wang
Yan
Y
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.
Gao
Yi
Y
Sun
Er-wei
EW
Xie
Jin-min
JM
Zhang
Hui-ying
HY
Chen
Jian-bo
JB
D019264:Adoptive Transfer / Q000379:methods; D000818:Animals; D017209:Apoptosis; D003907:Dexamethasone / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D005452:Fluoresceins / Q000737:chemistry* / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000737:chemistry / Q000493:pharmacokinetics; D008214:Lymphocytes / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D015203:Reproducibility of Results; D013154:Spleen / Q000166:cytology; D013388:Succinimides / Q000737:chemistry* / Q000493:pharmacokinetics
C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D005456:Fluorescent Dyes; D013388:Succinimides; D003907:Dexamethasone