Objective lt;br gt;To investigate the feasibility of labeling iris pigment epithelial(IPE)cells of rabbits with 5(and 6)carboxyfluorescein diacetate succinimidyl ester(CFSE). lt;br gt; lt;br gt;Methods lt;br gt;Enzyme-assisted microdissection was used to isolate the cultured rabbit prime;s IPE cells.The third or forth subcultured IPE cells were incubated with 2.5,5,10,20,and 40 mu;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. lt;br gt;Results lt;br gt;Incubation with 20 mu;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. lt;br gt; lt;br gt;Conclusion lt;br gt;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 prime;s IPE cells. lt;br gt; lt;br gt;(Chin J Ocul Fundus Dis, 2006, 22: 261-264)
Citation: PENG Shaomin,ZHANG Zhongyu,LI Xiaoxin,et al.. Iris pigment epithelial cells of rabbits stained by carboxyfluorescein diacetate succinimidyl ester in vitro. Chinese Journal of Ocular Fundus Diseases, 2006, 22(4): 261-264. doi: Copy