Limits...
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.

Show MeSH

Related in: MedlinePlus

In vitro expansion modulates VEGF-induced EPC migration VEGF-induced EPC chemotaxis was tested in a Boyden chamber migration assay. Cell expansion increased late EPC migration towards VEGF (10 ng/ml). Data are the numbers of migrating EPCs. The mean and SEM of three experiments are shown (*P= 0.023).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4401281&req=5

fig05: In vitro expansion modulates VEGF-induced EPC migration VEGF-induced EPC chemotaxis was tested in a Boyden chamber migration assay. Cell expansion increased late EPC migration towards VEGF (10 ng/ml). Data are the numbers of migrating EPCs. The mean and SEM of three experiments are shown (*P= 0.023).

Mentions: The functional implications of the increase in VEGFR2 density were then investigated in a Boyden chamber migration test with VEGF as chemo-attrac-tant in the lower chamber. We performed previous experiments with increasing VEGF doses (10, 50 and 100 ng/ml). VEGF induced a dose-dependent migration that was statistically significant for all concentrations used (data not shown). However, using 50 and 100 ng/ml of VEGF, the migrating cells were very numerous and the 10 ng/ml VEGF concentration was further chosen to better evaluate any difference in the migration potential. In these conditions, a significant increase in the chemotactic effect of VEGF was observed with EPCs expanded for 5 weeks compared to EPCs expanded for 3 weeks (P= 0.023, Fig. 5).


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)

In vitro expansion modulates VEGF-induced EPC migration VEGF-induced EPC chemotaxis was tested in a Boyden chamber migration assay. Cell expansion increased late EPC migration towards VEGF (10 ng/ml). Data are the numbers of migrating EPCs. The mean and SEM of three experiments are shown (*P= 0.023).
© Copyright Policy
Related In: Results  -  Collection

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

fig05: In vitro expansion modulates VEGF-induced EPC migration VEGF-induced EPC chemotaxis was tested in a Boyden chamber migration assay. Cell expansion increased late EPC migration towards VEGF (10 ng/ml). Data are the numbers of migrating EPCs. The mean and SEM of three experiments are shown (*P= 0.023).
Mentions: The functional implications of the increase in VEGFR2 density were then investigated in a Boyden chamber migration test with VEGF as chemo-attrac-tant in the lower chamber. We performed previous experiments with increasing VEGF doses (10, 50 and 100 ng/ml). VEGF induced a dose-dependent migration that was statistically significant for all concentrations used (data not shown). However, using 50 and 100 ng/ml of VEGF, the migrating cells were very numerous and the 10 ng/ml VEGF concentration was further chosen to better evaluate any difference in the migration potential. In these conditions, a significant increase in the chemotactic effect of VEGF was observed with EPCs expanded for 5 weeks compared to EPCs expanded for 3 weeks (P= 0.023, Fig. 5).

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