A Method of Isolating Endothelial Progenitor Cells from Mice

Objective To investigate an appropriate method for isolating EPCs from mice bone marrow.Methods The mononuclear cells from mice bone marrow collected by density gradient separation were cultured in M199 medium after examination by flow cytometry.After 7 days' culture,cells were stained with DIL-acLDL and FITC-UEA-1,and tested the abilities of migration and incorporation into the vascular network.Results The spindle cells grew dominantly after 7 days' culture.About 0.029%±0.008% of total mononuclear cells detected by flow cytometry were CD34+CD133+VEGFR2+ cells.The percentage of cells labeled both with DIL-acLDL and FITC-UEA-1 was 86.085%±5.622%.The number of migratory cells reached 19.458±2.251/field of view.The average cell number incorporating into the vascular network was 67.750±8.823/field of view detected by cell migration test.Conclusion The above method is a relatively ideal one for isolating and assessing EPCs from mice bone marrow,which could offer a better support to further research on EPCs.