Volume 64, Issue 5, Pages (November 2003)

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Volume 64, Issue 5, Pages 1648-1652 (November 2003) Endothelial progenitor cell proliferation and differentiation is regulated by erythropoietin Rapid Communication Ferdinand H. Bahlmann, Kirsten Degroot, Thorsten Duckert, Eva Niemczyk, Elisabeth Bahlmann, Sascha M. Boehm, Hermann Haller, Danilo Fliser Kidney International Volume 64, Issue 5, Pages 1648-1652 (November 2003) DOI: 10.1046/j.1523-1755.2003.00279.x Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 1 The number of circulating stem cells (cSCs) in whole blood of eight renal patients before and during darbepoetin treatment. #P < 0.05 comparison between baseline and week 6. Kidney International 2003 64, 1648-1652DOI: (10.1046/j.1523-1755.2003.00279.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 2 Quantitative assessment of cultured endothelial progenitor cells (EPCs) from eight renal patients during darbepoetin therapy. Administration of darbepoetin clearly resulted in a marked increase in total EPC number within 6 weeks of therapy. #P < 0.01 comparison of week 2 and week 6 vs. baseline. Kidney International 2003 64, 1648-1652DOI: (10.1046/j.1523-1755.2003.00279.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 3 Representative photomicrograph of cultured endothelial progenitor cells (EPCs) from a patient with renal anemia before and after 6 weeks of darbepoietin therapy. EPCs were stained with both Ulex europaeus agglutinin-1 (UEA-1) fluorescein isothiocyanate (FITC) (green fluorescence) and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine-labeled acetylated low-density lipoprotein (acLDL-DiI) (red fluorescence). Kidney International 2003 64, 1648-1652DOI: (10.1046/j.1523-1755.2003.00279.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 4 Dose-dependent effect of darbepoetin alfa on endothelial progenitor cells (EPCs). (A) Phase contrast images demonstrating the effect of darbepoetin on formation of tube-like structures by co-cultured EPCs in an in vitro angiogenesis assay. (B) Representative photomicrographs of tube formation with 0.005 μg/mL darbepoetin. Fluorescent-labeled EPCs (red) were co-plated with human umbilical vein endothelial cells (HUVECs) (transparent) to form tubular structures. Both cell types were stained with endothelial cell-specific Ulex europaeus agglutinin-1 (UEA-1) (green fluorescence). Superimposed phase-contrast and fluorescent images of identical fields reveal that EPCs made a substantial contribution to the tubular network. (C) Quantitative assessment of the tube formation index. #P < 0.05 vs. 0 μg/mL. Kidney International 2003 64, 1648-1652DOI: (10.1046/j.1523-1755.2003.00279.x) Copyright © 2003 International Society of Nephrology Terms and Conditions

Figure 5 Representative Western immunoblots of Akt phosphorylation are shown as time-dependent changes in Akt phosphorylation at serine 473 after exposure of endothelial progenitor cells (EPCs) to darbepoietin (0.005 μg/mL). p-Akt, phospho Akt (active protein kinase B); Akt 1, total Akt (active and inactive protein kinase B). Kidney International 2003 64, 1648-1652DOI: (10.1046/j.1523-1755.2003.00279.x) Copyright © 2003 International Society of Nephrology Terms and Conditions