Mature endothelium and neurons are simultaneously derived from embryonic stem cells by 2D in vitro culture system

Alessio Noghero, Marco Arese, Federico Bussolino, Anna Gualandris

Research output: Contribution to journalArticlepeer-review

Abstract

The connections existing between vessels and nerves go beyond the structural architecture of vascular and nervous systems to comprise cell fate determination. The analysis of functional/molecular links that interconnect endothelial and neural commitments requires a model in which the two differentiation programs take place at the same time in an artificial controllable environment. To this regard, this work presents an in vitro model to differentiate embryonic stem (ES) cells simultaneously into mature neurons and endothelial cells. Murine ES cells are differentiated within an artificial environment composed of PA6 stromal cells and a serum-free medium. Upon these basal culture conditions ES cells preferentially differentiate into neurons. The addition of basic fibroblast growth factor (FGF2) to the medium allows the simultaneous maturation of neurons and endothelial cells, whereas bone morphogenetic protein (BMP)4 drives endothelial differentiation to the disadvantage of neural commitment. The responsiveness of the system to exogenous cytokines was confirmed by genes expression analysis that revealed a significant up-regulation of endothelial genes in presence of FGF2 and a massive down-regulation of the neural markers in response to BMP4. Furthermore, the role played by single genes in determining endothelial and neural fate can be easily explored by knocking down the expression of the target gene with lentiviruses carrying the corresponding shRNA sequence. The possibility to address the neural and the endothelial fate separately or simultaneously by exogenous stimuli combined with an efficient gene silencing strategy make this model an optimal tool to identify environmental signals and genes pathways involved in both endothelial and neural specification.

Original languageEnglish
Pages (from-to)2200-2215
Number of pages16
JournalJournal of Cellular and Molecular Medicine
Volume15
Issue number10
DOIs
Publication statusPublished - Oct 2011

Keywords

  • Embryonic stem cells
  • Endothelial differentiation
  • Gene targeting
  • Neural differentiation
  • ShRNA
  • Stromal cells

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Medicine

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