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Isolation and characterization of undifferentiated Endothelial Cell Progenitors from murine adipose tissue

Isolation and characterization of undifferentiated Endothelial Cell Progenitors from murine adipose tissue

FASEB Journal 18(4-5): Abst 144 3

Recent studies have documented the presence of endothelial progenitor cells (EPC) in the bone marrow and peripheral blood. Fat tissue stroma has been shown to contain adult stem cells. We hypothesize that a considerable portion of adult stem cells are undifferentiated progenitors of vascular cells. To test this, murine adipose tissue was processed by collagenase digestion and differential centrifugation to obtain multipotent stromal cells. By fluorescent flow cell sorting we collected the DiI-acetyl-LDL-labeled (D+) mononuclear cells which accounts for a fraction (3+/-0.2%) of the total stromal cells in the primary cultures. When cultured in the presence of vascular endothelial growth factor (50 ng/mL VEGF), D+ cells generated cell colonies displaying an endothelial cell morphology. Flow cytometry, immunofluorescence and western blott revealed that the majority of D+ cells exhibited higher levels of Ulex europaeus agglutinin-1, VEGF-R1/2, CD34, CD133 and Tie-2 and lower levels of markers for mature endothelial cells such as eNOS and von Willebrand factor. Flow cytometry and immunofluorescence detected expression of the catalytic subunit of telomerase and cyclin D1 in these cells. These data illustrate the existence of vascular stem cells in the adipose tissue. The percentage of EPC appears higher than that in the bone marrow suggesting the fat tissue may serve as a valuable source of EPC for heart cell therapy. .

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