Bone marrow-derived endothelial progenitor cells isolation and culture

Objective To isolate and culture endothelial progenitor cells(EPCs).Methods Bone marrow cell were cultured in dishes coated with 1% gelatin with M199 medium supplemented with 10% fetal bovine serum and 50μg/mL endothelial cell growth supplements,at 37°C in a humidified atmosphere of 5% CO2.Subcultured using trypsin-EDTA.EPCs were identified by immunofluorescence staining for CD31 expression,ability of lectin binding.Further characterisation of EPCs was performed by analysing capillary tube formation.Results EPCs colonies appeared between 5 and 7 days of culture and defined as a central core of rounded cells.Two weeks later,cells presented the characteristic cobblestone morphology.The differentiation status of EPCs was confirmed by binding UEA-1 plant lectin and expressing CD31.Further,tubular network was formed.Conclusions When induced in culture,EPCs can be obtained,exhibit long term proliferative potential.