Endothelial Progenitor Cells Derived from Rat Bone Marrow Culture with Differential Adhesion Method

Objective:To explore an economic, reliable, stable and high purity of endothelial progenitor cells(EPCs) in culture methods. Methods:Rat bone marrow-derived EPCs were separated by density gradient centrifugation and cultured,which were then plated on 25 mL culture flask adherent culture with human fibronection. The no-adherent cells cultured for 48 h were collected and cultured for 7 days, and then identified by being observed the experiment of uptaking DiL-acLDL and combining FITC-UEA-1 by fluorescence microscope. Flow Cytometry was performed to analyze the expressions of CD133, flk-1, VE-cadherin (CD144) and CD31 dynamically. Results:Cells cultured 48 h without adherent were small and spindle or polygonal in shape. Large numbers of typical EPCs coloy-forming unites were found. Cells up-took DiI-acLDL and combined FITC-UEA-1 with fluorescent staining positive rate(87.0±6.0)%. The positive rates of cell surface antigen CD133, flk-1, CD144 and CD31 were 61.01%, 8.55%, 6.18% and 8.20% detected by flow cytometry. Conclusion:The differential attachment method is a reliable, stable and high purity of endothelial progenitor cell culture method.