Ascorbic acid improves pluripotency of human parthenogenetic embryonic stem cells through modifying imprinted gene expression in the Dlk1-Dio3 region

Stem Cell Res Ther. 2015 Apr 14;6(1):69. doi: 10.1186/s13287-015-0054-9.

Abstract

Introduction: Human parthenogenetic embryonic stem cells (hpESCs) are generated from artificially activated oocytes, however, the issue of whether hpESCs have equivalent differentiation ability to human fertilized embryonic stem cells remains controversial.

Methods: hpESCs were injected into male severe combined immunodeficiency (SCID) mice and the efficiency of teratoma formation was calculated. Then the gene expression and methylation modification were detected by real time-PCR and bisulfate methods.

Results: Comparison of five hpESCs with different differentiation abilities revealed that levels of paternal genes in the Dlk1-Dio3 region on chromosome 14 in the hpESCs with high differentiation potential are enhanced, but strictly methylated and silenced in the hpESCs with lower differentiation potential. Treatment with ascorbic acid, rescued their ability to support teratoma formation and altered the expression profiles of paternally expressed genes in hpESCs that could not form teratoma easily. No differences in the expression of other imprinting genes were evident between hpESCs with higher and lower differentiation potential, except for those in the Dlk1-Dio3 region.

Conclusions: The Dlk1-Dio3 imprinting gene cluster distinguishes the differentiation ability of hpESCs. Moreover, modification by ascorbic acid may facilitate application of hpESCs to clinical settings in the future by enhancing their pluripotency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ascorbic Acid / pharmacology*
  • Calcium-Binding Proteins
  • Cell Differentiation
  • DNA Methylation / genetics
  • Embryo Culture Techniques
  • Embryonic Stem Cells / drug effects*
  • Gene Expression / genetics
  • Gene Expression Profiling
  • Humans
  • Intercellular Signaling Peptides and Proteins / biosynthesis
  • Intercellular Signaling Peptides and Proteins / genetics*
  • Iodide Peroxidase / biosynthesis
  • Iodide Peroxidase / genetics*
  • Male
  • Membrane Proteins / biosynthesis
  • Membrane Proteins / genetics*
  • Mice
  • Mice, SCID
  • Multigene Family / genetics
  • Parthenogenesis
  • Pluripotent Stem Cells / drug effects
  • Teratoma / genetics
  • Teratoma / prevention & control*

Substances

  • Calcium-Binding Proteins
  • DLK1 protein, human
  • Intercellular Signaling Peptides and Proteins
  • Membrane Proteins
  • iodothyronine deiodinase type III
  • Iodide Peroxidase
  • Ascorbic Acid