Diverse Epigenetic Profile of Novel Human Embryonic Stem Cell Lines

Abstract

Previously published online as a Cell Cycle E-publication: http://www.landesbioscience.com/journals/cc/abstract.php?id=2440 KEY WORDShESCs, pluripotency related genes, epigenetic background, methylation ACKNOWLEDGEMENTSWe are very grateful to Dr.Suzanne Kadereit for critical reading the manuscript and helpful discussions. Report Diverse Epigenetic Profile of Novel Human Embryonic Stem Cell Lines ABSTRACTHuman embryonic stem cells (hESCs) are a promising model for studying mechanisms of regulation of early development and differentiation. OCT4, NANOG, OCT4-related genes and some others were recently described to be important in pluripotency maintenance. Lesser is known about molecular mechanisms involved in their regulation. Apart from genetic regulation of gene expression epigenetic events, particularly methylation, play an important role in early development. Using RT-PCR we studied the expression of pluripotency-related genes OCT4, NANOG, DPPA3 and DPPA5 during hESCs differentiation to embryoid bodies. Analysis of methylation profiles of promoter or putative regulatory regions of the indicated genes demonstrated that expression of the pluripotency-maintaining genes correlated with their methylation status, whereas methylation of DPPA3 and DPPA5 varied between cell lines. We propose that DNA methylation underlies the developmental stage-specific mechanisms of pluripotency-related genes expression and reactivation and may have an impact on differentiation potential of hESC lines.

A role for polyamine regulators in ESC self-renewal

“…Tianyun Zhao, 1 Kim Jee Goh, 2 Huck Hui Ng 2,3,4,5,6 and Leah A. Vardy 1,3, * 1 A*STAR Institute of Medical Biology; Singapore; 2 Gene Regulation Laboratory; Genome Institute of Singapore; Singapore; 3 School of Biological Sciences; Nanyang Technological University; Singapore; 4 Department of Biochemistry; Yong Loo Lin School of Medicine; National University of Singapore; Singapore; 5 Department of Biological Sciences; National University of Singapore; Singapore; 6 NUS Graduate School for Integrative Sciences and Engineering; National University of Singapore; Singapore of somatic cell reprogramming to produce induced pluripotent stem cells (iPSCs) has enabled the generation of disease and patient-specific iPSCs for disease modeling and potentially for therapeutic application. 1,2 Recently, significant progress has been made in understanding the molecular networks that regulate the decision of ESCs and iPSCs to self-renew or differentiate to specific lineages.…”

“…DNA methylation is dramatically altered on differentiation, and the promoters of pluripotency genes OCT4 and NANOG show reduced methylation in ESCs. 5 The polycomb repressive complex 2 (PRC2) is a histone-modifying complex that functions to repress transcription through histone methylation. Inactivation of the PRC complex proteins results in an enhanced ESC state and compromised differentiation, highlighting the essential role these complexes play in ESCs.…”

Isolation and characterization of stem cells derived from human third molar tooth germs of young adults: implications in neo-vascularization, osteo-, adipo- and neurogenesis

“…ESC messenger RNA (mRNA) from human embryonic stem cell line Moscow 01 (hESM01) was kindly provided by Dr Sergey L Kiselev (Institute of Gene Biology, Russian Academy of Sciences, Moscow). 32 Complementary DNA synthesis was conducted using random hexamer primers and moloney murine leukemia virus reverse transcriptase (MMLV-RT, Promega) at 37 1C for 1 h. For real-time PCR, primer sets of each gene and PCR conditions were chosen and applied as reported in the literature (Table 1). In addition, TaqMan primers and probes (Table 2) were designed using PrimerExpress (Applied Biosystems, Foster City, CA, USA) software.…”

An integrative analysis of reprogramming in human isogenic system identified a clone selection criterion

“…1) by introducing reprogramming factors into the previously described hESC line hESM01. 18,19 The cell line hESM01-OSKMN-DOX-n5 (hereafter referred to as n5) that expressed all transgenes exclusively in the presence of DOX in undifferentiated or differentiated states, had a normal karyotype and demonstrated pluripotency in vitro and in vivo was selected for further analysis (Fig. S1 and Supplemental Experimental Procedures).…”