Establishment of the method for stem cells labeling and tracking of umbilical cord-mesenchymal stem cells in BALB/c nude mice
Objective To establish the method for labeling and tracing umbilical cord mesenchymal stem cells (UC-MSCs) in vivo and to investigate the distribution of UC-MSCs in BALB/c nude mice.Methods DiR stain was used to label the UC-MSCs. The effects of DiR labeling on the viability,morphology,surface markers and cell cycle of UC-MSCs were evaluated. On this basis,the DiR labeled UC-MSCs were injected into BALB/c nude mice by tail vein at a dose of 3 × 106 cells. The distribution of UC-MSCs in BALB/c nude mice were detected by in vivo imaging and immunofluorescence.Results The optimal concentration and labeling time for DiR was 5 μg/ml and 5 min,respectively. There were no significant changes in cell morphology,cell activity,surface markers and cell cycle. After injection,UC-MSCs were rapidly distributed in the lung and liver,and subsequently in the spleen and other organs. UC-MSCs could not be detected by the living imaging and immunofluorescence at 30 days after transplantation.Conclusion DiR stain can be used to label UC-MSCs and has no significant effect on the cell biological characteristics of UC-MSCs. UC-MSCs cannot be widely distributed and remain for a long time in vivo after transplantation in mice,suggesting that UC-MSCs has a good preclinical safety.
mesenchymal stem cellsin vivo tracer techniqueimagingbiodistributionnonclinical research