Cytogenetic Analysis and Dlk1-Dio3 Locus Epigenetic Status of Mouse Embryonic Stem Cells During Early Passages

Overview

Journal Cytotechnology

Specialties Biotechnology
Genetics

Date 2014 Jun 28

PMID 24969018

Citations 3

Authors

Aleksei Menzorov

Inna Pristyazhnyuk

Helen Kizilova

Anastasia Yunusova

Nariman Battulin

Antonina Zhelezova

Aleftina Golubitsa

Oleg Serov

Affiliations

Soon will be listed here.

Abstract

Mouse embryonic stem (ES) cells are widely used in early development studies and for transgenic animal production; however, a stable karyotype is a prerequisite for their use. We derived 32 ES cell lines of outbred mice (129 × BALB (1B), C57BL × 1B, and DD × 1B F1 hybrids). Pluripotency was assessed by utilizing stem-cell-marker gene expression, teratoma formation assays and the formation of chimeras. It was shown that only 21 of the 32 ES cell lines had a diploid modal number of chromosomes of 40. In these lines, the percentage of diploid cells varied from 30.3 to 78.9 %, and trisomy of chromosomes 1, 8 and 11 was observed in some cells in 16.7, 36.7 and 20.0 % of the diploid ES cell lines, respectively. Some cells had trisomy of chromosomes 6, 9, 12, 14, 18 and 19. In situ hybridization with an X chromosome paint probe revealed that 7 of the 11 XX-cell lines had X chromosome rearrangements in some cells. Analysis of the methylation status of the Dlk1-Dio3 locus showed that imprinting was altered in 4 of the 18 ES cell lines. Thus, mouse ES cell lines are prone to chromosome abnormalities even at early passages. Therefore, routine cytogenetic and imprinting analyses are necessary for ES cell characterization.

Citing Articles

Mitochondria structural reorganization during mouse embryonic stem cell derivation.

Suldina L, Morozova K, Menzorov A, Kizilova E, Kiseleva E Protoplasma. 2018; 255(5):1373-1386.

PMID: 29549502 DOI: 10.1007/s00709-018-1236-y.

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.

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

Yu Y, Gao Q, Zhao H, Li R, Gao J, Ding T Stem Cell Res Ther. 2015; 6:69.

PMID: 25879223 PMC: 4425892. DOI: 10.1186/s13287-015-0054-9.

References

Park J, Yoshida I, Tada T, Takagi N, Takahashi Y, Kanagawa H . Differentiative potential of a mouse parthenogenetic embryonic stem cell line revealed by embryoid body formation in vitro. Jpn J Vet Res. 1998; 46(1):19-28. View

Ben-David U, Benvenisty N . High prevalence of evolutionarily conserved and species-specific genomic aberrations in mouse pluripotent stem cells. Stem Cells. 2012; 30(4):612-22. DOI: 10.1002/stem.1057. View

Stadtfeld M, Apostolou E, Akutsu H, Fukuda A, Follett P, Natesan S . Aberrant silencing of imprinted genes on chromosome 12qF1 in mouse induced pluripotent stem cells. Nature. 2010; 465(7295):175-81. PMC: 3987905. DOI: 10.1038/nature09017. View

Rebuzzini P, Neri T, Zuccotti M, Redi C, Garagna S . Chromosome number variation in three mouse embryonic stem cell lines during culture. Cytotechnology. 2008; 58(1):17-23. PMC: 2593760. DOI: 10.1007/s10616-008-9164-x. View

Eggan K, Akutsu H, Loring J, Klemm M, Rideout 3rd W, Yanagimachi R . Hybrid vigor, fetal overgrowth, and viability of mice derived by nuclear cloning and tetraploid embryo complementation. Proc Natl Acad Sci U S A. 2001; 98(11):6209-14. PMC: 33447. DOI: 10.1073/pnas.101118898. View

Cockburn K, Rossant J . Making the blastocyst: lessons from the mouse. J Clin Invest. 2010; 120(4):995-1003. PMC: 2846056. DOI: 10.1172/JCI41229. View

Takahashi K, Okita K, Nakagawa M, Yamanaka S . Induction of pluripotent stem cells from fibroblast cultures. Nat Protoc. 2007; 2(12):3081-9. DOI: 10.1038/nprot.2007.418. View

Sugawara A, Goto K, Sotomaru Y, Sofuni T, Ito T . Current status of chromosomal abnormalities in mouse embryonic stem cell lines used in Japan. Comp Med. 2006; 56(1):31-4. View

Liu X, Wu H, Loring J, Hormuzdi S, Disteche C, Bornstein P . Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission. Dev Dyn. 1997; 209(1):85-91. DOI: 10.1002/(SICI)1097-0177(199705)209:1<85::AID-AJA8>3.0.CO;2-T. View

10.

Martin G . Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci U S A. 1981; 78(12):7634-8. PMC: 349323. DOI: 10.1073/pnas.78.12.7634. View

11.

Pristiazhniuk I, Matveeva N, Grafodatskii A, Serdiukova N, Serov O . [Analysis of chromosome composition in interspecific embryonic stem hybrid cells of mice]. Tsitologiia. 2010; 52(2):136-43. View

12.

Rebuzzini P, Neri T, Mazzini G, Zuccotti M, Redi C, Garagna S . Karyotype analysis of the euploid cell population of a mouse embryonic stem cell line revealed a high incidence of chromosome abnormalities that varied during culture. Cytogenet Genome Res. 2008; 121(1):18-24. DOI: 10.1159/000124377. View

13.

Mantel C, Guo Y, Lee M, Kim M, Han M, Shibayama H . Checkpoint-apoptosis uncoupling in human and mouse embryonic stem cells: a source of karyotpic instability. Blood. 2007; 109(10):4518-27. PMC: 1885509. DOI: 10.1182/blood-2006-10-054247. View

14.

Wang Z, Jaenisch R . At most three ES cells contribute to the somatic lineages of chimeric mice and of mice produced by ES-tetraploid complementation. Dev Biol. 2004; 275(1):192-201. DOI: 10.1016/j.ydbio.2004.06.026. View

15.

Choppa P, Vojdani A, Tagle C, Andrin R, Magtoto L . Multiplex PCR for the detection of Mycoplasma fermentans, M. hominis and M. penetrans in cell cultures and blood samples of patients with chronic fatigue syndrome. Mol Cell Probes. 1998; 12(5):301-8. DOI: 10.1006/mcpr.1998.0186. View

16.

Evans M, Kaufman M . Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981; 292(5819):154-6. DOI: 10.1038/292154a0. View

17.

Battulin N, Khabarova A, Boyarskikh U, Menzorov A, Filipenko M, Serov O . Reprogramming somatic cells by fusion with embryonic stem cells does not cause silencing of the Dlk1-Dio3 region in mice. World J Stem Cells. 2012; 4(8):87-93. PMC: 3506971. DOI: 10.4252/wjsc.v4.i8.87. View

18.

Bryja V, Bonilla S, cajanek L, Parish C, Schwartz C, Luo Y . An efficient method for the derivation of mouse embryonic stem cells. Stem Cells. 2005; 24(4):844-9. DOI: 10.1634/stemcells.2005-0444. View

19.

da Rocha S, Edwards C, Ito M, Ogata T, Ferguson-Smith A . Genomic imprinting at the mammalian Dlk1-Dio3 domain. Trends Genet. 2008; 24(6):306-16. DOI: 10.1016/j.tig.2008.03.011. View

20.

Nagy A, Rossant J, Nagy R, Abramow-Newerly W, Roder J . Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc Natl Acad Sci U S A. 1993; 90(18):8424-8. PMC: 47369. DOI: 10.1073/pnas.90.18.8424. View