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Human Molecular Genetics Advance Access published online on November 30, 2005
Human Molecular Genetics, doi:10.1093/hmg/ddi427
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© The Author 2005. Published by Oxford University Press. All rights reserved
Received September 22, 2005
Accepted November 12, 2005

Article

Temporal and Parental Specific Expression of Imprinted Genes in a Newly Derived Chinese Human Embryonic Stem Cell Line and Embryoid Bodies

Bo Wen Sun 1, A Cong Yang 2, Yun Feng 3, Yi Juan Sun 3, Yu fei Zhu 4, Yi Zhang 4, Hua Jiang 4, Chun Liang Li 4, Fu Rong Gao 4, Zhi Hong Zhang 4, Wei Cheng Wang 1, Xiang Yin Kong 4, Gang Jin 4, Shi Jun Fu 4, and Ying Jin 5 *

1 Institute of Health Science, Shanghai Institutes for Biological Sciences of CAS and Shanghai JiaoTong University School of Medicine, 225 Chongqing Nan Road, Shanghai, 200025, China; Department of Molecular Developmental Biology, Shanghai JiaoTong University School of Medicine, Shanghai, 200025, China
2 Department of Molecular Developmental Biology, Shanghai JiaoTong University School of Medicine, Shanghai, 200025, China; Institute of Health Science, Shanghai Institutes for Biological Sciences of CAS and Shanghai JiaoTong University School of Medicine, 225 Chongqing Nan Road, Shanghai, 200025, China
3 Reproductive Medical Center, Ruijin Hospital, Shanghai, China
4 Institute of Health Science, Shanghai Institutes for Biological Sciences of CAS and Shanghai JiaoTong University School of Medicine, 225 Chongqing Nan Road, Shanghai, 200025, China
5 Institute of Health Science, Shanghai Institutes for Biological Sciences of CAS and Shanghai JiaoTong University School of Medicine, Room 607, Building 1, 225 Chongqing Nan Road, Shanghai, 200025, China; Department of Molecular Developmental Biology, Shanghai JiaoTong University School of Medicine, Shanghai, 200025, China

* To whom correspondence should be addressed.
Ying Jin, E-mail: yjin{at}sibs.ac.cn


  Abstract

Although the study of imprinted genes in human development is very important, little is known about their expression and regulation in the early differentiation of human tissues due to lack of an appropriate model. In this study, a Chinese human embryonic stem (hES) cell line, SHhES1, was derived and fully characterized. Expression profiles of human imprinted genes were determined by Affymetrix Oligo micro-array in undifferentiated SHhES1 cells and SHhES1 derived embryoid bodies (EBs) at day -3, 8, 13, and 18. Thirty-two known human imprinted genes were detected in undifferentiated ES cells. Significantly, differential expression was found in nine genes at different stages of EB formation. Expression profile changes were confirmed by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) in SHhES1 cells as well as in another independently derived hES cell line, HUES-7. In addition, the mono-allelic expressions of four imprinted genes were examined in three different passages of undifferentiated ES cells and EBs of both hES cell lines. The mono-allelic expressions of imprinted genes, H19, PEG10, NDNL1, and KCNQ1 were maintained in both undifferentiated hES cells and derived EBs. More importantly, with availability of maternal peripheral blood lymphocyte (PBL) sample, we demonstrated that the maternal expression of KCNQ1 and paternal expression of NDNL1 and PEG10 were maintained in SHhES1 cells. These data provide the first demonstration that parental specific expression of imprinted genes is stable in EBs after extensive differentiation, also indicating that in vitro fertilization protocol does not disrupt the parental mono-allelic expression of the imprinted genes examined.


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