Mesenchymal Stem Cells of Rats Stained by Carboxyfluorescein Diacetate Succinimidy Esterin vitro

Objective To investigate the feasibility of labeling mesenchymal stem cells(MSCs)of rats with 5,6,2-carboxyfluorescein diacetate succinimidy ester(CFSE)and the optimal condition.Methods MSCs obtained from 20 SD rats,bone marrow were isolated,purified and cultured in vitro with Percoll density gradient centrifugation combined with adherent method.MSCs of the third generation MSCs filling in the floor of 25cm2 cultured bottles were incubated with 2.5,5,10,20,and 40μmol/L of CFSE for 1,5,10,15,and 20 min respectively.The optimal concentration and labeling time of CFSE were indentified by detecting the fluorescence intensity and adhesive rate of labeled MSCs.The proliferative activity of MSCs labeled by CFSE under the fittest condition was tested by MTT method.Results Incubation with 10μmol/L of CFSE under 37℃ for 10 minutes was the optimal condition for MSCs labeling.The labeled MSCs maintained the same proliferative ability as the non-labeled MSCs under the optimal condition.Conclusion With the advantages of high rate of dyeing,low cell toxicity,safety and convenience,CFSE can be used as a new method to label the rat's MSC.Incubation with 10 μmol/L of CFSE under 37℃ for 10 minutes is the optimal condition for MSCs labeling.