Biological characteristics of endothelial progenitor cells derived from granulocyte colony-stimulating factor mobilized peripheral blood / 中国组织工程研究

ABSTRACT

BACKGROUND:Endothelial progenitor cells have good prospects for clinical application;especial y as seed cells, they are involved in construction of tissue engineered blood vessels and disease treatment. OBJECTIVE:To investigate the biological characteristics of endothelial progenitor cells derived from granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood. METHODS:Mononuclear cells from G-CSF mobilized peripheral blood (n=9) and normal adult peripheral blood (n=8) were obtained and cultured in expanded medium. The immunophenotype of endothelial progenitor cells was investigated by FACS and immunohistochemistry. Real-time PCR was used to detect the expression of different cytokines. Proliferation and adhesion ability of endothelial progenitor cells were detected by MTT assay and adhesion assay. Moreover, the abilities of vasculogenesis in vitro and Dil-labeled acetylated low density lipoprotein uptake were detected. The acquired cells were seeded onto the basilar membrane gel containing vascular endothelial growth factor and basic fibroblast growth factor to induce angiogenesis. RESULTS AND CONCLUSION:Endothelial progenitor cells derived from G-CSF mobilized peripheral blood were similar to those from normal adult peripheral blood in phenotype and morphology. FACSs and immunohistochemistry showed that endothelial progenitor cells were positive for endothelial cellmarkers, such as CD31, vWF, CD34, FLK-1, VE-Cadherin and CD133. In addition, the expression of vascular endothelial growth factor and stromal cel-derived factor 1 were significantly increased in G-CSF mobilized endothelial progenitor cells. Moreover, endothelial progenitor cells derived from two sources possessed the abilities of angiogenesis in vitro and Dil-labeled acetylated low density lipoprotein uptake. But, the number and the proliferation ability of endothelial progenitor cells derived from G-CSF mobilized peripheral blood were better than that of endothelial progenitor cells derived from normal adult peripheral blood. These findings indicate that there is a population of cells with characteristics of endothelial progenitor cells in G-CSF mobilized peripheral blood. They possess the abilities of vasculogenesis in vitro. Moreover, compared to endothelial progenitor cells derived from normal adult peripheral blood, we could obtain more endothelial progenitor cells in G-CSF mobilized peripheral blood and those cells show better proliferation ability.