In vitro culture and biological characterization of rat bone marrow-derived endothelial progenitor cells

Objective To evaluate the characterization of rat bone marrowderived endothelial progenitor cell(EPC) cultured and induced in vitro.Methods Mononuclear cells were collected from bone marrow suspension by density gradient centrifugation.The growth of EPCs induced with and without bFGF were compared.The cells were identified by immunocytochemistry.The CD133+/Flk-1+ markers were checked by flow cytometer on day 3,7 and 14.Results The adherent cells were able to line up in the typical cord-like structure and formed clusters.Adherent cells showed CD133,CD34,Flk-1,Ⅷ/ vWF positive.Flow cytometry analysis indicated that the number of CD133+/Flk-1+ double positive cells in the induced group were significant higher than that in the non-induced group(P0.01).Conclusion EPC could be isolated and cultured by density gradient centrifugation and subjected to culture with bFGF induction.These cells show the characterization of endothelial cells,and could differentiate into endothelial cells.