Tip Vascular Endothelial Cells. Galectin-3 Regulation Of The Mechanism Of The Effect Of Plastic Pipe

Part I Isolation and differentiation of human endothelial progen-itor cells and Tip vascular endothelial cells[Abjective] To investigate the methods of isolation and cultivation of human endothelial progenitor cells (EPCs). To Research the culture conditions and biological characteristics of human endothelial progenitor cells in vitro. To confirm the peripheral blood is the another idear source of human endothelial progenitor cells except the bone marrow. The human endothelial progenitor cells were differentiated to Tip vascular endothelial cells. This can provide the cells for next experiment and a reliable basis for angiogeneisis.[Methods] Peripheral blood mononuclear cells were isolated by density-gradient centrifugation and incubated in endothelial medium in the presence of vascular endothelial growth factor. Observe the morphology of the cells every day. The cells were detached at day7, and the indentified by endothelial-specific markers. The expression of CD34,CD133and VEGFR were determinded by flow cytometry. After21days, the vWF and eNOS were determinded by reverse transcriptase-polymerase chain reaction(RT-PCR).[Results] Attached spindle-like cells appeared after4days culture and become more and large after7days. The positive rates of CD34,CD133and VEGFR of ex vivo expanded endothelial progenitor cells were (68.9±2.34)%,(81.7±1.50)%,(97.1±2.01)%. After21days, the pebble-like cells spread the filopodia and connect each other. RT-PCR demonstrated the presence of vWF and eNOS mRNA.[Conclusion] Endothelial progenitor cells could be successfully isolated and cultured from perpheral blood mononuclear cells. The human endothelial progenitor cells also were differentiated to Tip vascular endothelial cells. Part IIEffects of Galectin-3on proliferation of Tip vascular endothelial cell derived from peripheral blood endothelial progenitor cell[Abjective] Endothelial progenitor cells is the precursor of the endothelial cells. The cells can transfer to peripheral blood from bone marrow and participate in the vascular repair and angiogenesis. Galectin-3is a galactose binding protein and play a major role in angiogen-esis. We observe the effects of Galectin-3on proliferation of Tip vascular endothelial cell derived from peripheral blood endothelial progenitor cell. This could provied more evidences for treating lower limb ischemia by stem cells.[Methods] The cultured peripheral blood endothelial progenitor cell in vitro were isolated and purified from human peripheral blood, and the cells were differentiated into Tip vascular endothelial cells. After21days, the cells were cultivated with the galectin-3for24hours with concentrations of0ug/ml,0.1ug/ml,1.0ug/ml,2.5ug/ml,5.0ug/ml,10ug/ml. The proliferation of Tip vascular endothelial cells derived from peripheral blood endothelial progenitor cell was measured by MTT.[Results] There were significant differences on the OD of six concentrations group (F=37.329, P<0.01). The OD of the concentrations group (5.0ug/ml and10.Oug/ml) of galectin-3were higher than the concentrations group (Oug/ml,0.1ug/ml,1.Oug/ml and2.5ug/ml), P<0.05. There were significant differences on the OD between the5.0ug/ml group and10.0ug/ml group (0.887±0.193vs1.154±0.303, P<0.05). There were positive correlation betwe-en the proliferation effects of Tip vascular endothelial cell derived from peripheral blood endothelial progenitor cell and the concentrations of Galectin-3(r=0.966)[Conclusion] Galectin-3can promote the proliferation of Tip vascular endothelial cells derived from peripheral blood endothelial progenitor cell.