Fibronectin promotes VEGF-induced CD34+ cell differentiation into endothelial cells Errol S Wijelath, PhD, Salman Rahman, PhD, Jacqueline Murray, PhD,

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Fibronectin promotes VEGF-induced CD34+ cell differentiation into endothelial cells Errol S Wijelath, PhD, Salman Rahman, PhD, Jacqueline Murray, PhD, Yatin Patel, PhD, Geoffrey Savidge, MD, Michael Sobel, MD Journal of Vascular Surgery Volume 39, Issue 3, Pages 655-660 (March 2004) DOI: 10.1016/j.jvs.2003.10.042

Fig 1 Comparison of ECM proteins on VEGF-induced CD34+ differentiation. 3 × 105 CD34+ cells were cultured for 21 days with endothelial growth medium on ECM-VEGF–coated plates. The number of endothelial colonies formed was identified by staining for vWF and VE-cadherin. Results are expressed as mean ± SD (n = 3). #P < .05 compared with control (15% serum). Journal of Vascular Surgery 2004 39, 655-660DOI: (10.1016/j.jvs.2003.10.042)

Fig 2 RT-PCR analysis for endothelial markers from VEGF-FN–differentiated CD34+ cells. A, RNA was extracted from freshly isolated CD34+ cells (day 0) and VEGF-FN differentiated CD34+ cells (day 21) and was analyzed for endothelial-specific markers by RT-PCR. Data shown are representative of two independent experiments. B, Densitometry analysis of endothelial markers. Data are expressed as ratio of mRNA levels to gel background. Data are means ± SEM (n = 2). Journal of Vascular Surgery 2004 39, 655-660DOI: (10.1016/j.jvs.2003.10.042)

Fig 3 Effect of ECM proteins on VEGF-induced CD34+ cell migration. Migration assays were carried out as described in the Methods section. The migration rates of CD34+ cells to VEGF and different combinations of ECM proteins were measured after 6 hours. Results are expressed as mean ± SD (n = 3). #P < .05 compared with VEGF or FN alone. Negative control was medium alone, and positive controls were VEGF or FN. Journal of Vascular Surgery 2004 39, 655-660DOI: (10.1016/j.jvs.2003.10.042)

Fig 4 Inhibition of VEGF-FN–induced CD34+ cell differentiation and migration by antibodies to α5β1. A, CD34+ cells were cultured on FN-VEGF–coated plates in endothelial growth medium containing blocking antibodies for α5β1, for αvβ3, or for αvβ5 (5 μg/mL). Fresh antibodies were added every 3 days. Endothelial colonies were determined after 21 days by staining for vWF and VE-cadherin. Results are expressed as mean ± SD (n = 3). B, Blocking antibodies to α5β1, αvβ3, or αvβ5 (20 μg/mL) were assessed for inhibition of VEGF-FN–induced CD34+ cell migration. Results are expressed as mean ± SD (n = 3). #P < .05 compared with VEGF-FN. Journal of Vascular Surgery 2004 39, 655-660DOI: (10.1016/j.jvs.2003.10.042)