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Mol. Cells 2009; 27(6): 635-640

Published online June 30, 2009

https://doi.org/10.1007/s10059-009-0084-1

© The Korean Society for Molecular and Cellular Biology

H19 Gene Is Epigenetically Stable in MouseMultipotent Germline Stem Cells

Shin Hye Oh, Yoon Hee Jung, Mukesh Kumar Gupta, Sang Jun Uhm, and Hoon Taek Lee

Received: December 10, 2009; Revised: April 14, 2009; Accepted: April 21, 2009

Abstract

Testis-derived germline stem (GS) cells can undergo re-programming to acquire multipotency when cultured under appropriate culture conditions. These multipotent GS (mGS) cells have been known to differ from GS cells in their DNA methylation pattern. In this study, we examined the DNA methylation status of the H19 imprinting control region (ICR) in multipotent adult germline stem (maGS) cells to elucidate how epigenetic imprints are altered by culture conditions. DNA methylation was analyzed by bisulfite sequencing PCR of established maGS cells cultured in the presence of glial cell line-derived neurotrophic factor (GDNF) alone or both GDNF and leukemia inhibitory factor (LIF). The results showed that the H19 ICR in maGS cells of both groups was hypermethylated and had an androgenetic pattern similar to that of GS cells. In line with these data, the relative abundance of the Igf2 mRNA transcript was two-fold higher and that of H19 was three fold lower than in control embryonic stem cells. The androgenetic DNA methylation pattern of the H19 ICR was maintained even after 54 passages. Furthermore, differentiating maGS cells from retinoic acid-treated embryoid bodies main-tained the androgenetic imprinting pattern of the H19 ICR. Taken together these data suggest that our maGS cells are epigenetically stable for the H19 gene during in vitro modifications. Further studies on the epigenetic regulation and chromatin structure of maGS cells are therefore necessary before their full potential can be utilized in regenerative medicine.

Keywords DNA methylation, genomic imprinting, germline stem cells, H19, Igf2

Article

Research Article

Mol. Cells 2009; 27(6): 635-640

Published online June 30, 2009 https://doi.org/10.1007/s10059-009-0084-1

Copyright © The Korean Society for Molecular and Cellular Biology.

H19 Gene Is Epigenetically Stable in MouseMultipotent Germline Stem Cells

Shin Hye Oh, Yoon Hee Jung, Mukesh Kumar Gupta, Sang Jun Uhm, and Hoon Taek Lee

Received: December 10, 2009; Revised: April 14, 2009; Accepted: April 21, 2009

Abstract

Testis-derived germline stem (GS) cells can undergo re-programming to acquire multipotency when cultured under appropriate culture conditions. These multipotent GS (mGS) cells have been known to differ from GS cells in their DNA methylation pattern. In this study, we examined the DNA methylation status of the H19 imprinting control region (ICR) in multipotent adult germline stem (maGS) cells to elucidate how epigenetic imprints are altered by culture conditions. DNA methylation was analyzed by bisulfite sequencing PCR of established maGS cells cultured in the presence of glial cell line-derived neurotrophic factor (GDNF) alone or both GDNF and leukemia inhibitory factor (LIF). The results showed that the H19 ICR in maGS cells of both groups was hypermethylated and had an androgenetic pattern similar to that of GS cells. In line with these data, the relative abundance of the Igf2 mRNA transcript was two-fold higher and that of H19 was three fold lower than in control embryonic stem cells. The androgenetic DNA methylation pattern of the H19 ICR was maintained even after 54 passages. Furthermore, differentiating maGS cells from retinoic acid-treated embryoid bodies main-tained the androgenetic imprinting pattern of the H19 ICR. Taken together these data suggest that our maGS cells are epigenetically stable for the H19 gene during in vitro modifications. Further studies on the epigenetic regulation and chromatin structure of maGS cells are therefore necessary before their full potential can be utilized in regenerative medicine.

Keywords: DNA methylation, genomic imprinting, germline stem cells, H19, Igf2

Mol. Cells
Nov 30, 2022 Vol.45 No.11, pp. 763~867
COVER PICTURE
Naive (cyan) and axotomized (magenta) retinal ganglion cell axons in Xenopus tropicalis (Choi et al., pp. 846-854).

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