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

Yang Yu,Hong-Cui Zhao,Jiang-Man Gao,Qian Gao,Rong Li,Jie Qiao,Si-Yu Bao,Xiao-Fang Sun,Ting Ding,Yong Fan,Yue Zhao

doi:10.1186/s13287-015-0054-9

Abstract

IntroductionHuman 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.MethodshpESCs 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.ResultsComparison 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.ConclusionsThe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0054-9) contains supplementary material, which is available to authorized users.

Highlights

Human parthenogenetic embryonic stem cells are generated from artificially activated oocytes, the issue of whether hpESCs have equivalent differentiation ability to human fertilized embryonic stem cells remains controversial
We examined the effects of ascorbic acid on methylation and gene expression patterns in the Dlk1Dio3 cluster, with a view to ascertaining whether this compound contributes to improvement of the differentiation ability of hpESCs
Data from the present study suggest that the differences in gene expression and methylation modifications in the Dlk1-Dio3 region contribute to the differentiation ability of hpESCs

Summary

Introduction

Human parthenogenetic embryonic stem cells (hpESCs) are generated from artificially activated oocytes, the issue of whether hpESCs have equivalent differentiation ability to human fertilized embryonic stem cells remains controversial. Human embryonic stem cells (hESCs) have tremendous potential in regenerative medicine and cell therapy, as they can differentiate into cell derivatives of all three primary germ layers, endoderm, mesoderm and ectoderm. Application of established cloned hESCs to the clinic is difficult at present because the hemagglutinating virus of Japan is pECSs have been derived from mouse [4], rabbit [5], monkey [6] and human sources [7,8,9,10], and their ability to differentiate into neurons and endoderm cells has been widely confirmed. Defects in differentiation ability of hpESCs limit their clinical application

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Conclusion