Reprogramming Somatic Cells by Fusion with Embryonic Stem Cells Does Not Cause Silencing of the Dlk1-Dio3 Region in Mice

Overview

Journal World J Stem Cells

Publisher Baishideng Publishing Group

Date 2012 Nov 29

PMID 23189213

Citations 5

Authors

Nariman R Battulin

Anna A Khabarova

Ulyana A Boyarskikh

Aleksey G Menzorov

Maxim L Filipenko

Oleg L Serov

Affiliations

Soon will be listed here.

Abstract

Aim: To examine the imprinted Dlk1-Dio3 locus in pluripotent embryonic stem (ES) cell/fibroblast hybrid cells.

Methods: Gtl2, Rian, and Mirg mRNA expression in mouse pluripotent ES cell/fibroblast hybrid cells was examined by real-time reverse transcription-polymerase chain reaction. Pyrosequencing and bisulfate sequencing were used to determine the DNA methylation level of the Dlk1-Dio3 locus imprinting control region.

Results: The selected hybrid clones had a near-tetraploid karyotype and were highly pluripotent judging from their capacity to generate chimeric embryos and adult chimeras. Our data clearly demonstrate that Gtl2, Rian, and Mirg, which are imprinted genes within the Dlk1-Dio3 locus, are active in all examined ES cell/fibroblast hybrid clones. In spite of interclonal variability, the expression of the imprinted genes is comparable to that of ES cells and fibroblasts. Quantitative analysis of the DNA methylation status of the intergenic differentially methylated region (IG DMR) within the Dlk1-Dio3 locus by pyrosequencing and bisulfite sequencing clearly showed that the DNA methylation status of the imprinted region in the tested hybrid clones was comparable to that of both ES cells and fibroblasts.

Conclusion: Reprogramming process in a hybrid cell system is achieved without marked alteration of the imprinted Dlk1-Dio3 locus.

Citing Articles

Embryonic germ cell extracts erase imprinted genes and improve the efficiency of induced pluripotent stem cells.

Hu J, Zhao Q, Feng Y, Li N, Gu Y, Sun R Sci Rep. 2018; 8(1):10955.

PMID: 30026469 PMC: 6053380. DOI: 10.1038/s41598-018-29339-0.

Alternative dominance of the parental genomes in hybrid cells generated through the fusion of mouse embryonic stem cells with fibroblasts.

Matveeva N, Fishman V, Zakharova I, Shevchenko A, Pristyazhnyuk I, Menzorov A Sci Rep. 2017; 7(1):18094.

PMID: 29273752 PMC: 5741742. DOI: 10.1038/s41598-017-18352-4.

Serum DLK1 is a potential prognostic biomarker in patients with hepatocellular carcinoma.

Li H, Cui M, Chen T, Xie H, Cui Y, Tu H Tumour Biol. 2015; 36(11):8399-404.

PMID: 26018510 DOI: 10.1007/s13277-015-3607-8.

Cytogenetic analysis and Dlk1-Dio3 locus epigenetic status of mouse embryonic stem cells during early passages.

Menzorov A, Pristyazhnyuk I, Kizilova H, Yunusova A, Battulin N, Zhelezova A Cytotechnology. 2014; 68(1):61-71.

PMID: 24969018 PMC: 4698258. DOI: 10.1007/s10616-014-9751-y.

Oct4 promoter activity in stem cells obtained through somatic reprogramming.

Krueger W, Tanasijevic B, Norris C, Tian X, Rasmussen T Cell Reprogram. 2013; 15(2):151-8.

PMID: 23550731 PMC: 3616451. DOI: 10.1089/cell.2012.0059.

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