Abstract:

BACKGROUND: A safe, efficient, and convenient labeling method with high sensitivity and strong specificity is needed to understand the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro and in vivo.
OBJECTIVE: To investigate the efficacy of labeling BMSCs with 5-ethynyl-2’-deoxyuridin (EdU) and its effects on cell proliferation.
METHODS: BMSCs were isolated by adherence method from bone marrow of rats and cultured to the third generation. BMSCs were seeded into 96-well plate with concentration of 5×107/L. BMSCs were labeled with EdU at different concentrations (0, 10, 20 and 50 μmol/L). The EdU-labeling positive rates and morphology of BMSCs in each group were detected at 24, 48 and 72 hours of culture, and the cell growth curves were obtained.
RESULTS AND CONCLUSION: The EdU-labeling positive rates were gadually increased in each group as time went on (P < 0.05), and the differences in the increased EdU-labeling positive rates were not significant (P > 0.05). Good EdU-labeling positive rates could be obtained in the 10 μmol/L group at 72 hours of culture. The BMSCs growth curves of each group before and after labeling were “S”-shaped. This suggested that application of EdU labeling within the experimental concentrations for BMSCs in vitro is efficient and safe without effects on cell proliferation. Good labeling rate of BMSCs can be obtained through labeling with EdU in 10 μmol/L for 72 hours.