Human endothelial colony forming cells from adult peripheral blood have enhanced sprouting angiogenic potential through up-regulating VEGFR2 signaling

Hyung Joon Joo, Sukhyun Song, Ha Rim Seo, Jennifer H. Shin, Seung Cheol Choi, Jae Hyoung Park, Cheol Woong Yu, Soon Jun Hong, Do-Sun Lim

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Abstract Background: Endothelial colony forming cells (ECFCs), a subtype of endothelial progenitor cells, have been studied as a promising cellular source for therapeutic angiogenesis. Although ECFCs are very similar to mature endothelial cells, details regarding the role of ECFCs during angiogenesis are not known. We compared the cellular and angiogenic properties of ECFCs and mature endothelial cells (HUVECs). Methods: HUVECs were used as control. Quantitative RT-PCR, western blotting, immunofluorescence staining, flow cytometric analyses and angiogenic cytokine array were performed. 3D-microfluidic angiogenesis assay system was adopted for in vitro angiogenic potential. In vivo angiogenic potential was assessed by Matrigel plug assay. Results: ECFCs had higher expression of activated endothelial tip cell markers (Dll4, CXCR4, CD34, and VCAM1) and arterial genes (DLL4 and CX40), but lower expression of venous and lymphatic genes (COUP-TFII and PROX1). In 3D-microfluidic angiogenesis assay system, ECFCs induced robust sprouting vascular structures. Co-cultivation of both ECFCs and HUVECs gave rise to lumen-formed hybrid vascular structures, with the resulting ECFCs predominantly localized to the tip portion. This finding suggests that the ECFC has a role as a sprouting endothelial tip cell. Interestingly, VEGF-A phosphorylated VEGFR2 and its downstream signaling molecules more strongly in ECFCs than in HUVECs. Even small amount of VEGF-A successfully induced the sprouting angiogenesis of ECFCs. Finally, co-administration of ECFCs and human dermal fibroblasts successfully induced lumen-formed maturated neovessels in vivo. Conclusion: ECFCs derived from adult peripheral blood had enhanced sprouting angiogenic potential in vitro and in vivo through up-regulation of the VEGFR2 signaling pathway.

Original languageEnglish
Article number20669
Pages (from-to)33-43
Number of pages11
JournalInternational Journal of Cardiology
Volume197
DOIs
Publication statusPublished - 2015 Aug 5

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Endothelial Cells
Microfluidics
Human Umbilical Vein Endothelial Cells
Vascular Endothelial Growth Factor A
Blood Vessels
Genes
Fluorescent Antibody Technique
Up-Regulation
Fibroblasts
Western Blotting
Staining and Labeling
Cytokines
Polymerase Chain Reaction
Skin
In Vitro Techniques
Therapeutics

Keywords

  • Angiogenesis
  • Endothelial progenitor cell
  • Microfluidic system
  • Vascular endothelial growth factor receptor

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Human endothelial colony forming cells from adult peripheral blood have enhanced sprouting angiogenic potential through up-regulating VEGFR2 signaling. / Joo, Hyung Joon; Song, Sukhyun; Seo, Ha Rim; Shin, Jennifer H.; Choi, Seung Cheol; Park, Jae Hyoung; Yu, Cheol Woong; Hong, Soon Jun; Lim, Do-Sun.

In: International Journal of Cardiology, Vol. 197, 20669, 05.08.2015, p. 33-43.

Research output: Contribution to journalArticle

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abstract = "Abstract Background: Endothelial colony forming cells (ECFCs), a subtype of endothelial progenitor cells, have been studied as a promising cellular source for therapeutic angiogenesis. Although ECFCs are very similar to mature endothelial cells, details regarding the role of ECFCs during angiogenesis are not known. We compared the cellular and angiogenic properties of ECFCs and mature endothelial cells (HUVECs). Methods: HUVECs were used as control. Quantitative RT-PCR, western blotting, immunofluorescence staining, flow cytometric analyses and angiogenic cytokine array were performed. 3D-microfluidic angiogenesis assay system was adopted for in vitro angiogenic potential. In vivo angiogenic potential was assessed by Matrigel plug assay. Results: ECFCs had higher expression of activated endothelial tip cell markers (Dll4, CXCR4, CD34, and VCAM1) and arterial genes (DLL4 and CX40), but lower expression of venous and lymphatic genes (COUP-TFII and PROX1). In 3D-microfluidic angiogenesis assay system, ECFCs induced robust sprouting vascular structures. Co-cultivation of both ECFCs and HUVECs gave rise to lumen-formed hybrid vascular structures, with the resulting ECFCs predominantly localized to the tip portion. This finding suggests that the ECFC has a role as a sprouting endothelial tip cell. Interestingly, VEGF-A phosphorylated VEGFR2 and its downstream signaling molecules more strongly in ECFCs than in HUVECs. Even small amount of VEGF-A successfully induced the sprouting angiogenesis of ECFCs. Finally, co-administration of ECFCs and human dermal fibroblasts successfully induced lumen-formed maturated neovessels in vivo. Conclusion: ECFCs derived from adult peripheral blood had enhanced sprouting angiogenic potential in vitro and in vivo through up-regulation of the VEGFR2 signaling pathway.",
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author = "Joo, {Hyung Joon} and Sukhyun Song and Seo, {Ha Rim} and Shin, {Jennifer H.} and Choi, {Seung Cheol} and Park, {Jae Hyoung} and Yu, {Cheol Woong} and Hong, {Soon Jun} and Do-Sun Lim",
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T1 - Human endothelial colony forming cells from adult peripheral blood have enhanced sprouting angiogenic potential through up-regulating VEGFR2 signaling

AU - Joo, Hyung Joon

AU - Song, Sukhyun

AU - Seo, Ha Rim

AU - Shin, Jennifer H.

AU - Choi, Seung Cheol

AU - Park, Jae Hyoung

AU - Yu, Cheol Woong

AU - Hong, Soon Jun

AU - Lim, Do-Sun

PY - 2015/8/5

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N2 - Abstract Background: Endothelial colony forming cells (ECFCs), a subtype of endothelial progenitor cells, have been studied as a promising cellular source for therapeutic angiogenesis. Although ECFCs are very similar to mature endothelial cells, details regarding the role of ECFCs during angiogenesis are not known. We compared the cellular and angiogenic properties of ECFCs and mature endothelial cells (HUVECs). Methods: HUVECs were used as control. Quantitative RT-PCR, western blotting, immunofluorescence staining, flow cytometric analyses and angiogenic cytokine array were performed. 3D-microfluidic angiogenesis assay system was adopted for in vitro angiogenic potential. In vivo angiogenic potential was assessed by Matrigel plug assay. Results: ECFCs had higher expression of activated endothelial tip cell markers (Dll4, CXCR4, CD34, and VCAM1) and arterial genes (DLL4 and CX40), but lower expression of venous and lymphatic genes (COUP-TFII and PROX1). In 3D-microfluidic angiogenesis assay system, ECFCs induced robust sprouting vascular structures. Co-cultivation of both ECFCs and HUVECs gave rise to lumen-formed hybrid vascular structures, with the resulting ECFCs predominantly localized to the tip portion. This finding suggests that the ECFC has a role as a sprouting endothelial tip cell. Interestingly, VEGF-A phosphorylated VEGFR2 and its downstream signaling molecules more strongly in ECFCs than in HUVECs. Even small amount of VEGF-A successfully induced the sprouting angiogenesis of ECFCs. Finally, co-administration of ECFCs and human dermal fibroblasts successfully induced lumen-formed maturated neovessels in vivo. Conclusion: ECFCs derived from adult peripheral blood had enhanced sprouting angiogenic potential in vitro and in vivo through up-regulation of the VEGFR2 signaling pathway.

AB - Abstract Background: Endothelial colony forming cells (ECFCs), a subtype of endothelial progenitor cells, have been studied as a promising cellular source for therapeutic angiogenesis. Although ECFCs are very similar to mature endothelial cells, details regarding the role of ECFCs during angiogenesis are not known. We compared the cellular and angiogenic properties of ECFCs and mature endothelial cells (HUVECs). Methods: HUVECs were used as control. Quantitative RT-PCR, western blotting, immunofluorescence staining, flow cytometric analyses and angiogenic cytokine array were performed. 3D-microfluidic angiogenesis assay system was adopted for in vitro angiogenic potential. In vivo angiogenic potential was assessed by Matrigel plug assay. Results: ECFCs had higher expression of activated endothelial tip cell markers (Dll4, CXCR4, CD34, and VCAM1) and arterial genes (DLL4 and CX40), but lower expression of venous and lymphatic genes (COUP-TFII and PROX1). In 3D-microfluidic angiogenesis assay system, ECFCs induced robust sprouting vascular structures. Co-cultivation of both ECFCs and HUVECs gave rise to lumen-formed hybrid vascular structures, with the resulting ECFCs predominantly localized to the tip portion. This finding suggests that the ECFC has a role as a sprouting endothelial tip cell. Interestingly, VEGF-A phosphorylated VEGFR2 and its downstream signaling molecules more strongly in ECFCs than in HUVECs. Even small amount of VEGF-A successfully induced the sprouting angiogenesis of ECFCs. Finally, co-administration of ECFCs and human dermal fibroblasts successfully induced lumen-formed maturated neovessels in vivo. Conclusion: ECFCs derived from adult peripheral blood had enhanced sprouting angiogenic potential in vitro and in vivo through up-regulation of the VEGFR2 signaling pathway.

KW - Angiogenesis

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KW - Microfluidic system

KW - Vascular endothelial growth factor receptor

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