Establishment of sprouting embryoid body model mimicking early embryonic vasculogenesis in human embryo

Hua Jiang, You Ji Feng, Yi Xie, Jin Lan Han, Zack Z. Wang, Tong Chen

Research output: Contribution to journalArticle

Abstract

Objective: To establish a sprouting embryoid body model mimicking early embryonic vasculogenesis in human embryo. Methods: Human embryonic stem were (hESCs) were cultured on the mouse embryo fibloblasts and then were induced to differentiate to form three-dimensional EB. The hEBs were cultured in media containing various angiogenesis-related factors: vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), endostatin, angiostatin, and platelet factor (PF)-4 of different concentrations for 3 days to observe the sprouting of the hEBs. 3, 3, 3′, 3′-tetramethylindo-carbocyanine perchlorate labeled acetylated low density lipoprotein (Dil-AcLDL) was added onto the hEBs foe 4 h Immunofluorescence assay was used to observe if Dil-AcLDL was absorbed and if CD31 was expressed so as to determine the existence of embryonic endothelial cells in the sprouting structures. The ideal culturing condition was analyzed. Results: The differentiated EBs formed sprouting structures in the collagen I matrix containing VEGF and FGF. The sprouts among individual EBs were able to link to each other and form vascular network-like structures. In the presence of VEGF and FGF, the sprouts branching from the EBs assimilated Dil-AcLDL, expressed CD31 and formed a 3-dimensional cylindrical organization. The concentrations of growth factors ideally stimulating sprouting growth were 100 ng/ml of VEGF and 50 ng/ml of FGF. The networks among the EBs were abolished by the angiostatin, endostatin, and PF4. Conclusion: The sprouting from hEBs accumulates embryonic endothelial cells and the sprouting network-like structures are indeed endothelial in nature. Inducing of sprouting EBs is an ideal model that mimics early embryonic vasculogenesis in humans.

Original languageEnglish (US)
Pages (from-to)2647-2651
Number of pages5
JournalZhonghua yi xue za zhi
Volume88
Issue number37
Publication statusPublished - Oct 14 2008
Externally publishedYes

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Keywords

  • Differentiation
  • Embryo body
  • Embryo development
  • Embryonic stem cell, human
  • Vasculogenesis

ASJC Scopus subject areas

  • Medicine(all)

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