不同浓度rhEPO对大鼠内皮祖细胞体外培养的影响

OBJECTIVE: Some researches demonstrate that recombinant human erythropoietin (rhEPO) can mobilize Endothelial progenitor cells (EPCs) from bone marrow. To investigate the effect of different concentration rhEPO on the function of rats EPCs. METHODS: Bone marrow was obtained from both femurs and tibials of Wistar rats, and mononuclear cells were isolated by density gradient centrifugation, followed by cultured and treated with several does of rhEPO (0, 4, 8 U/mL). At 7 days after culture, EPCs were identified by FITC-UEA-1 and Dil-Ac-LDL co-staining. Meanwhile, EPCs were marked by CD133 and CD34, identified by flow cytometry; additionally, the functions of EPCs were evaluated using adhesion test, migration assay and proliferation experiment. RESULTS AND CONCLUSION: At 7 days after culture, the cell morphology changed completely, and formed a tube-like structure, which presented typical cobblestone appearance after cell conjugation. The fluorescence staining showed that the EPCs were positive to both FITC-UEA-1 and Dil-Ac-LDL co-staining. The CD133 and CD34 cells were combined to EPCs by flow cytometry. Finally, the adhesion test, migration assay and proliferation experiment demonstrated that rhEPO could improve adhesion, migration and proliferative capacities in a dose-dependent manner. The results demonstrated that rhEPO can significantly enhance the functions of EPCs in dose-dependent manner at 0-8 U/mL concentration.