Efficient differentiation of hESCs into endothelial cells in vitro is secured by epigenetic changes

Cell Cycle. 2008 Sep 15;7(18):2929-35. doi: 10.4161/cc.7.18.6700.

Abstract

Human embryonic stem cells (hESCs) are to be considered as a valuable source for regenerative medicine because of their capacity to differentiate into all cell types. We have developed an efficient culture system to differentiate hECSs into endothelial cells without the formation of embryoid bodies Establishing appropriate culture conditions with a cocktail of growth factors allowed us to differentiate hESCs directly to endothelial primary culture with about 50% efficiency. CD31 immunomagnetic cell sorting was used to purify derived endothelium from the primary culture of hESCs. Isolated endothelial cells expressed immunological markers (vWF, CD105), specific genes (VE-cadherin, KDR, GATA-2, GATA-3, eNOS), and formed cord-like structures on collagen matrix and in Matrigel assay. During differentiation to endothelial lineage promoter regions of the genes involved in specific cell fate determination and homeostasis (GATA-2,-3, and eNOS) underwent intensive hypomethylation which correlated with the gene expression. Overall our data demonstrate that direct differentiation of hESCs leads to endothelial cells that acquire epigenetic patterning similar to the functional endothelial cells of the organism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Differentiation*
  • Cell Line
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / enzymology
  • Embryonic Stem Cells / metabolism*
  • Endothelial Cells / cytology*
  • Endothelial Cells / metabolism
  • Epigenesis, Genetic*
  • Gene Expression Regulation, Developmental
  • Humans
  • Immunomagnetic Separation
  • Mice
  • Nitric Oxide Synthase Type III / genetics
  • Nitric Oxide Synthase Type III / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA

Substances

  • Biomarkers
  • Nitric Oxide Synthase Type III