Iris pigment epithelial cells of rabbits stained by carboxyfluorescein diacetate succinimidyl ester in vitro
- VernacularTitle:活体染料羧基荧光素乙酰乙酸对兔眼虹膜色素上皮细胞体外染色的研究
- Author:
Shaomin PENG
;
Zhongyu ZHANG
;
Xiaoxin LI
- Publication Type:Journal Article
- Keywords:
Fluorescent dyes;
Iris;
Pigment epithelium of eye;
Staining and labeling
- From:
Chinese Journal of Ocular Fundus Diseases
2000;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the feasibility of labeling iris pigment epithelial(IPE)cells of rabbits with 5-(and 6-)carboxyfluorescein diacetate succinimidyl ester(CFSE). Methods Enzyme-assisted microdissection was used to isolate the cultured rabbit′s IPE cells.The third or forth subcultured IPE cells were incubated with 2.5,5,10,20,and 40 ?mol/L of CFSE for 1,5,and10min respectively.The fluorescence intensity was detected by flow cytometry,and the leakage of CFSE and its dyeing were observed by fluorescence antibody labeling. Results Incubation with 20 ?mol/L CFSE under 37℃for1minute was the most optimal condition for IPE cells labeling.The coloration of IPE cells stained by CFSE lasted 4 weeks.There was no leakage of dye from labeled rabbit IPE cells to non-labeled human IPE cells in mixed culture process. Conclusion With the advantages of high rate of dyeing,long time of tracing,safety and convenience,CFSE can be used as a new method to label the rabbit′s IPE cells.
