Culture and identification of endothelial progenitor cells from human adipose tissue with CD31 immunomagnetic beads / 中国组织工程研究

ABSTRACT

BACKGROUND:Endothelial progenitor cells are precursor cells of mature endothelial cells,which can migrate to ischemic tissues and differentiate into mature endothelial cells,and then play an important role in vascular remodeling.Endothelial progenitor cells have wide application prospects in various ischemic diseases,but the biological characteristics and identification methods are still controversial.OBJECTIVE:To investigate the methods of isolation and culture of endothelial progenitor cells from the human adipose tissue and to identify their biological features,in order to provide a sufficient source of cells for ischemic diseases.METHODS:Stromal vascular fraction cells were isolated from the human adipose tissue by enzymatic digestion,CD31+ cells were selected using immunomagnetic beads,and then cultured in endothelial basal medium-2 supplemented with the EGM-2-MV-SingleQuots.Endothelial progenitor cells were identified through detection of morphology,cell markers and cell functions.RESULTS AND CONCLUSION:(1) CD31 + cells were selected by immunomagnetic beads and then cultured and amplified in vitro,which displayed typical cobblestone-like morphology,and they maintain their proliferative ability.(2) Flow cytometry results showed that the CD31+ cells expressed CD31 (98.84％),CD34 (97.21％),VEGRR2 (64.07％),CD146 (98.42％) and CD133 (2.55％),but hardly expressed CD45 (1.1％),a hematopoietic stem cell marker.(3) The CD31 + cells were also found to incept Dil-ac-LDL and exhibit lectin binding capability.Furthermore,a lumen-like structure was formed in Matrigel,which has the ability of angiogenesis in vitro.To conclude,these results suggest that it is feasible to isolate and culture endothelial progenitor cells from the human adipose tissue by enzymatic digestion combined with immunomagnetic bead sorting.