Isolation,Culture and Identification of Bone Marrow Derived Endothelial Progenitor Cells

Objective:To explore the rat bone marrow derived endothelial progenitor cells(EPCs),culture,differentiation and identification method.Methods:The marrow cavity of rat femur was rinsed,single nuclear cells was obtained by density gradient centrifugation,those endothelial cells were cultured,the cell morphology was observed.CD34,vWF,CD133 were detected with immunohistochemistry and immunofluorescence,absorbed DiL-acLDL and combined with FITC Lectin-UEA-1,ultrastructure and chromosome karyotype analysis for identification.Results: The new separation of bone marrow mononuclear cells and rounded.After some cells began to stick wall,a spindle or irregular shape,7 d show cell colony or linear arrangement,10 d close to convergence,a typical cobblestone appearance.Adherent cells,CD34,vWF,CD133 positive expression and shows dynamic changes,could absorb Dil acLDL and combined with FITC Lectin UEA-1.Conclusion:This experiment preliminarily established a set of EPCs isolation,culture,differentiation of rat bone marrow.