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Increased VEGFR2 expression during human late endothelial progenitor cells expansion enhances in vitro angiogenesis with up-regulation of integrin alpha(6).

Smadja DM, Bièche I, Helley D, Laurendeau I, Simonin G, Muller L, Aiach M, Gaussem P - J. Cell. Mol. Med. (2007 Sep-Oct)

Bottom Line: Quantitative flow cytometry confirmed that VEGFR2 density on EPCs increased during the expansion process and was significantly higher than on HUVECs.VEGFR2 up-regulation had no effect on VEGF-induced cell proliferation, but significantly enhanced EPC migration and pseudotubes formation dependent on integrin alpha(6) subunit overexpression.In vitro expansion of late EPCs increases the expression of VEGFR2, the main VEGF receptor, with possible implications for EPC-based angiogenic therapy.

View Article: PubMed Central - PubMed

Affiliation: AP-HP, Service d'Hématologie Biologique A, Hôpital Européen Georges Pompidou, Paris, France.

ABSTRACT
In vitro expansion of late endothelial progenitor cells (EPCs) might yield a cell therapy product useful for myocardial and leg ischaemia, but the influence of EPC expansion on the angiogenic properties of these cells is unknown. In the present study, we investigated the effect of in vitro EPC expansion on vascular endothelial growth factor (VEGF) receptor expression. EPCs were obtained from CD34(+) cord blood cells and expanded for up to 5 weeks. Real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) showed that VEGFR2 expression, contrary to VEGFR1 and VEGFR3 expression, was significantly higher on expanded EPCs than on freshly isolated CD34(+) cells or on human umbilical vein endothelial cells (HUVECs). Quantitative flow cytometry confirmed that VEGFR2 density on EPCs increased during the expansion process and was significantly higher than on HUVECs. The impact of VEGFR2 increase was studied on the three theoretical steps of angiogenesis, i.e., EPC proliferation, migration and differentiation. VEGFR2 up-regulation had no effect on VEGF-induced cell proliferation, but significantly enhanced EPC migration and pseudotubes formation dependent on integrin alpha(6) subunit overexpression. In vitro expansion of late EPCs increases the expression of VEGFR2, the main VEGF receptor, with possible implications for EPC-based angiogenic therapy.

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VEGFR1, 2 and 3 and CD31 mRNA quantification in CD34+ cells, late EPCs, and HUVECs. mRNA levels were normalized to TBP mRNA levels and to the sample with the lowest quantifiable level (i.e. 1 on the left ordinate, corresponding to a Ct value of 35). Values above 100 represent a strong gene expression. Mean and SEM values of three different colonies are shown at each point.
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fig02: VEGFR1, 2 and 3 and CD31 mRNA quantification in CD34+ cells, late EPCs, and HUVECs. mRNA levels were normalized to TBP mRNA levels and to the sample with the lowest quantifiable level (i.e. 1 on the left ordinate, corresponding to a Ct value of 35). Values above 100 represent a strong gene expression. Mean and SEM values of three different colonies are shown at each point.

Mentions: Expression of VEGF receptor family genes was measured by real-time quantitative RT-PCR in EPCs expanded for 3 and 5 weeks, and also in freshly isolated CD34+ cells and HUVECs. The results are shown as ‘normalized mRNA levels’, with reference to the smallest quantifiable amount of mRNA defined by a Ct value of 35 (= 1 on the left log ordinate of Fig. 2; see Methods).


Increased VEGFR2 expression during human late endothelial progenitor cells expansion enhances in vitro angiogenesis with up-regulation of integrin alpha(6).

Smadja DM, Bièche I, Helley D, Laurendeau I, Simonin G, Muller L, Aiach M, Gaussem P - J. Cell. Mol. Med. (2007 Sep-Oct)

VEGFR1, 2 and 3 and CD31 mRNA quantification in CD34+ cells, late EPCs, and HUVECs. mRNA levels were normalized to TBP mRNA levels and to the sample with the lowest quantifiable level (i.e. 1 on the left ordinate, corresponding to a Ct value of 35). Values above 100 represent a strong gene expression. Mean and SEM values of three different colonies are shown at each point.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401281&req=5

fig02: VEGFR1, 2 and 3 and CD31 mRNA quantification in CD34+ cells, late EPCs, and HUVECs. mRNA levels were normalized to TBP mRNA levels and to the sample with the lowest quantifiable level (i.e. 1 on the left ordinate, corresponding to a Ct value of 35). Values above 100 represent a strong gene expression. Mean and SEM values of three different colonies are shown at each point.
Mentions: Expression of VEGF receptor family genes was measured by real-time quantitative RT-PCR in EPCs expanded for 3 and 5 weeks, and also in freshly isolated CD34+ cells and HUVECs. The results are shown as ‘normalized mRNA levels’, with reference to the smallest quantifiable amount of mRNA defined by a Ct value of 35 (= 1 on the left log ordinate of Fig. 2; see Methods).

Bottom Line: Quantitative flow cytometry confirmed that VEGFR2 density on EPCs increased during the expansion process and was significantly higher than on HUVECs.VEGFR2 up-regulation had no effect on VEGF-induced cell proliferation, but significantly enhanced EPC migration and pseudotubes formation dependent on integrin alpha(6) subunit overexpression.In vitro expansion of late EPCs increases the expression of VEGFR2, the main VEGF receptor, with possible implications for EPC-based angiogenic therapy.

View Article: PubMed Central - PubMed

Affiliation: AP-HP, Service d'Hématologie Biologique A, Hôpital Européen Georges Pompidou, Paris, France.

ABSTRACT
In vitro expansion of late endothelial progenitor cells (EPCs) might yield a cell therapy product useful for myocardial and leg ischaemia, but the influence of EPC expansion on the angiogenic properties of these cells is unknown. In the present study, we investigated the effect of in vitro EPC expansion on vascular endothelial growth factor (VEGF) receptor expression. EPCs were obtained from CD34(+) cord blood cells and expanded for up to 5 weeks. Real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) showed that VEGFR2 expression, contrary to VEGFR1 and VEGFR3 expression, was significantly higher on expanded EPCs than on freshly isolated CD34(+) cells or on human umbilical vein endothelial cells (HUVECs). Quantitative flow cytometry confirmed that VEGFR2 density on EPCs increased during the expansion process and was significantly higher than on HUVECs. The impact of VEGFR2 increase was studied on the three theoretical steps of angiogenesis, i.e., EPC proliferation, migration and differentiation. VEGFR2 up-regulation had no effect on VEGF-induced cell proliferation, but significantly enhanced EPC migration and pseudotubes formation dependent on integrin alpha(6) subunit overexpression. In vitro expansion of late EPCs increases the expression of VEGFR2, the main VEGF receptor, with possible implications for EPC-based angiogenic therapy.

Show MeSH
Related in: MedlinePlus