In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells

Zong jin Li, Chen Yang, Feng wu Tang, Zhi hua Zhang, Bin Xu, Qin jun Zhao, Ren chi Yang, Zack Z. Wang, Zhong chao Han

Research output: Contribution to journalArticle

Abstract

OBJECTIVE: To explore an optional condition to induce mouse embryonic stem (ES) cells to differentiate into endothelial cells and to establish in vitro models of vasculogenesis and angiogenesis. METHODS: Mouse ES cells were cultured in differentiation medium containing a cocktail of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), interleukin-6 (IL-6) and erythropoietin (EPO) in 1% methylcellulose to induce formation of embryoid bodies (EBs). At day 11, EBs were harvested and suspended in rat-tail collagen type I with the same cocktail of cytokines cultured for three additional days. The differentiation of ES cells into endothelial cells, processes of vasculogenesis and angiogenesis were examined using immunostaining of EBs slices and whole-mount immunocytochemistry of EBs with monoclonal antibodies (mAbs) against platelet endothelial cell adhesion molecule-1 (PECAM-1) and alpha-smooth muscle actin (SMA). RESULTS: Under appropriate culture conditions; ES cells spontaneously differentiated and formed EBs containing vascular structures and tubular channels, which were positive for PECAM-1 co-differentiated with smooth muscle. When not treated with angiogenic growth factors, PECAM-1-positive cells could not organize into vascular structures of 11-day-old EBs. In the presence of angiogenic factors 11-day old EBs embedded into type I collagen, and rapidly developed an endothelial networks. Whole-mount immunocytochemistry of collagen gel with anti-PECAM-1 antibody showed the formation of primary vascular structures sprouting from EBs. Quantitative analysis revealed that 100 microg/ml thalidomide significantly reduced the number and length of EBs endothelial sprouting. CONCLUSIONS: Mouse ES cells can differentiate into endothelial cells combined with smooth muscle differentiation during EBs formation and further develop endothelial outgrowths after EBs embedded into collagen, which respectively recapitulate vasculogenesis, angiogenesis, and arteriogenesis processes in vivo. The models provide a useful tool to investigate vasculogenesis, angiogenesis, and arteriogenesis mechanisms and evaluate the effects of angiogenic and angiostatic agents.

Original languageEnglish (US)
Pages (from-to)62-66
Number of pages5
JournalZhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae.
Volume27
Issue number1
StatePublished - Feb 2005
Externally publishedYes

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Embryoid Bodies
CD31 Antigens
Smooth Muscle
Blood Vessels
Endothelial Cells
Angiogenesis Inducing Agents
Embryonic Stem Cells
Collagen Type I
Fibroblast Growth Factor 6
In Vitro Techniques
Mouse Embryonic Stem Cells
Collagen
Immunohistochemistry
Methylcellulose
Thalidomide
Angiogenesis Inhibitors
Fibroblast Growth Factor 2
Erythropoietin
Vascular Endothelial Growth Factor A
Antibody Formation

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Li, Z. J., Yang, C., Tang, F. W., Zhang, Z. H., Xu, B., Zhao, Q. J., ... Han, Z. C. (2005). In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells. Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae., 27(1), 62-66.

In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells. / Li, Zong jin; Yang, Chen; Tang, Feng wu; Zhang, Zhi hua; Xu, Bin; Zhao, Qin jun; Yang, Ren chi; Wang, Zack Z.; Han, Zhong chao.

In: Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae., Vol. 27, No. 1, 02.2005, p. 62-66.

Research output: Contribution to journalArticle

Li, ZJ, Yang, C, Tang, FW, Zhang, ZH, Xu, B, Zhao, QJ, Yang, RC, Wang, ZZ & Han, ZC 2005, 'In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells', Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae., vol. 27, no. 1, pp. 62-66.
Li, Zong jin ; Yang, Chen ; Tang, Feng wu ; Zhang, Zhi hua ; Xu, Bin ; Zhao, Qin jun ; Yang, Ren chi ; Wang, Zack Z. ; Han, Zhong chao. / In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells. In: Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae. 2005 ; Vol. 27, No. 1. pp. 62-66.
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abstract = "OBJECTIVE: To explore an optional condition to induce mouse embryonic stem (ES) cells to differentiate into endothelial cells and to establish in vitro models of vasculogenesis and angiogenesis. METHODS: Mouse ES cells were cultured in differentiation medium containing a cocktail of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), interleukin-6 (IL-6) and erythropoietin (EPO) in 1{\%} methylcellulose to induce formation of embryoid bodies (EBs). At day 11, EBs were harvested and suspended in rat-tail collagen type I with the same cocktail of cytokines cultured for three additional days. The differentiation of ES cells into endothelial cells, processes of vasculogenesis and angiogenesis were examined using immunostaining of EBs slices and whole-mount immunocytochemistry of EBs with monoclonal antibodies (mAbs) against platelet endothelial cell adhesion molecule-1 (PECAM-1) and alpha-smooth muscle actin (SMA). RESULTS: Under appropriate culture conditions; ES cells spontaneously differentiated and formed EBs containing vascular structures and tubular channels, which were positive for PECAM-1 co-differentiated with smooth muscle. When not treated with angiogenic growth factors, PECAM-1-positive cells could not organize into vascular structures of 11-day-old EBs. In the presence of angiogenic factors 11-day old EBs embedded into type I collagen, and rapidly developed an endothelial networks. Whole-mount immunocytochemistry of collagen gel with anti-PECAM-1 antibody showed the formation of primary vascular structures sprouting from EBs. Quantitative analysis revealed that 100 microg/ml thalidomide significantly reduced the number and length of EBs endothelial sprouting. CONCLUSIONS: Mouse ES cells can differentiate into endothelial cells combined with smooth muscle differentiation during EBs formation and further develop endothelial outgrowths after EBs embedded into collagen, which respectively recapitulate vasculogenesis, angiogenesis, and arteriogenesis processes in vivo. The models provide a useful tool to investigate vasculogenesis, angiogenesis, and arteriogenesis mechanisms and evaluate the effects of angiogenic and angiostatic agents.",
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T1 - In vitro vasculogenesis and angiogenesis of mouse embryonic stem cells

AU - Li, Zong jin

AU - Yang, Chen

AU - Tang, Feng wu

AU - Zhang, Zhi hua

AU - Xu, Bin

AU - Zhao, Qin jun

AU - Yang, Ren chi

AU - Wang, Zack Z.

AU - Han, Zhong chao

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N2 - OBJECTIVE: To explore an optional condition to induce mouse embryonic stem (ES) cells to differentiate into endothelial cells and to establish in vitro models of vasculogenesis and angiogenesis. METHODS: Mouse ES cells were cultured in differentiation medium containing a cocktail of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), interleukin-6 (IL-6) and erythropoietin (EPO) in 1% methylcellulose to induce formation of embryoid bodies (EBs). At day 11, EBs were harvested and suspended in rat-tail collagen type I with the same cocktail of cytokines cultured for three additional days. The differentiation of ES cells into endothelial cells, processes of vasculogenesis and angiogenesis were examined using immunostaining of EBs slices and whole-mount immunocytochemistry of EBs with monoclonal antibodies (mAbs) against platelet endothelial cell adhesion molecule-1 (PECAM-1) and alpha-smooth muscle actin (SMA). RESULTS: Under appropriate culture conditions; ES cells spontaneously differentiated and formed EBs containing vascular structures and tubular channels, which were positive for PECAM-1 co-differentiated with smooth muscle. When not treated with angiogenic growth factors, PECAM-1-positive cells could not organize into vascular structures of 11-day-old EBs. In the presence of angiogenic factors 11-day old EBs embedded into type I collagen, and rapidly developed an endothelial networks. Whole-mount immunocytochemistry of collagen gel with anti-PECAM-1 antibody showed the formation of primary vascular structures sprouting from EBs. Quantitative analysis revealed that 100 microg/ml thalidomide significantly reduced the number and length of EBs endothelial sprouting. CONCLUSIONS: Mouse ES cells can differentiate into endothelial cells combined with smooth muscle differentiation during EBs formation and further develop endothelial outgrowths after EBs embedded into collagen, which respectively recapitulate vasculogenesis, angiogenesis, and arteriogenesis processes in vivo. The models provide a useful tool to investigate vasculogenesis, angiogenesis, and arteriogenesis mechanisms and evaluate the effects of angiogenic and angiostatic agents.

AB - OBJECTIVE: To explore an optional condition to induce mouse embryonic stem (ES) cells to differentiate into endothelial cells and to establish in vitro models of vasculogenesis and angiogenesis. METHODS: Mouse ES cells were cultured in differentiation medium containing a cocktail of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), interleukin-6 (IL-6) and erythropoietin (EPO) in 1% methylcellulose to induce formation of embryoid bodies (EBs). At day 11, EBs were harvested and suspended in rat-tail collagen type I with the same cocktail of cytokines cultured for three additional days. The differentiation of ES cells into endothelial cells, processes of vasculogenesis and angiogenesis were examined using immunostaining of EBs slices and whole-mount immunocytochemistry of EBs with monoclonal antibodies (mAbs) against platelet endothelial cell adhesion molecule-1 (PECAM-1) and alpha-smooth muscle actin (SMA). RESULTS: Under appropriate culture conditions; ES cells spontaneously differentiated and formed EBs containing vascular structures and tubular channels, which were positive for PECAM-1 co-differentiated with smooth muscle. When not treated with angiogenic growth factors, PECAM-1-positive cells could not organize into vascular structures of 11-day-old EBs. In the presence of angiogenic factors 11-day old EBs embedded into type I collagen, and rapidly developed an endothelial networks. Whole-mount immunocytochemistry of collagen gel with anti-PECAM-1 antibody showed the formation of primary vascular structures sprouting from EBs. Quantitative analysis revealed that 100 microg/ml thalidomide significantly reduced the number and length of EBs endothelial sprouting. CONCLUSIONS: Mouse ES cells can differentiate into endothelial cells combined with smooth muscle differentiation during EBs formation and further develop endothelial outgrowths after EBs embedded into collagen, which respectively recapitulate vasculogenesis, angiogenesis, and arteriogenesis processes in vivo. The models provide a useful tool to investigate vasculogenesis, angiogenesis, and arteriogenesis mechanisms and evaluate the effects of angiogenic and angiostatic agents.

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