Karyotype analysis of the euploid cell population of a mouse embryonic stem cell line revealed a high incidence of chromosome abnormalities that varied during cultureRebuzzini P.a · Neri T.a · Mazzini G.b · Zuccotti M.c · Redi C.A.a, d · Garagna S.a
aDipartimento di Biologia Animale, Laboratorio di Biologia dello Sviluppo, Università degli Studi di Pavia, bIstituto di Genetica Molecolare del CNR, Sezione di Istochimica e Citometria, Dipartimento di Biologia Animale, Pavia, cDipartimento di Medicina Sperimentale, Sezione di Istologia ed Embriologia, Università degli Studi di Parma, Parma, dFondazione IRCCS Policlinico San Matteo, Pavia (Italy)
Do you have an account?
- Rent for 48h to view
- Buy Cloud Access for unlimited viewing via different devices
- Synchronizing in the ReadCube Cloud
- Printing and saving restrictions apply
Rental: USD 8.50
Cloud: USD 20.00
It is common knowledge that mouse embryonic stem cell (mESC) lines accumulate chromosomal changes during culture. Despite the wide use of mESCs as a model of early mammalian development and cell differentiation, there is a lack of systematic studies aimed at characterizing their karyological changes during culture. We cultured an mESC line, derived in our laboratory, for a period of 3 months investigating its chromosome complement at different times. About 60% of the metaphases analysed were euploid throughout the culture period but, from passage 13, only 50% of the euploid metaphases had a proper chromosome complement. The remaining 50% showed chromosome abnormalities, mainly gain or loss of entire chromosomes, both within the same passage and among different passages analysed. The very heterogeneous spectrum of abnormalities indicates a high frequency of chromosome mutations that arise continuously during culture. The heterogeneity of the aberrant chromosome constitution of 2n = 40 metaphases, observed at different passages of culture, might be due either to their elimination or to a shift towards the hypoeu- or hypereuploid population of those metaphases that accumulate further chromosome abnormalities. The stability of the frequency of eu-, hypoeu- and hypereuploid populations during culture might, however, be due to the elimination of those cells that carry a high mutational burden. Based on our results, we suggest that karyotype analysis of the euploid cell population of mESC lines is necessary when such lines are used in the production of chimeric mice, for their contribution to the germ line, or when they are differentiated into specific cell types.
© 2008 S. Karger AG, Basel
- Aflatoonian B, Moore H: Germ cells from mouse and human embryonic stem cells. Reproduction 132:699–707 (2006).
- Brown T, Fox DP, Robertson FW, Bullock I: Non-random chromosome loss in PHA-stimulated lymphocytes from normal individuals. Mutat Res 122:403–406 (1983).
- Buzzard JJ, Gough NM, Crook JM, Colman A: Karyotype of human ES cells during extended culture. Nat Biotechnol 22:381–382 (2004).
- Caisander G, Park H, Frej K, Lindqvist J, Bergh C, et al: Chromosomal integrity maintained in five human embryonic stem cell lines after prolonged in vitro culture. Chromosome Res 14:131–137 (2006).
- Cowan CA, Klimanskaya I, McMahon J, Atienza J, Witmyer J, et al: Derivation of embryonic stem-cell lines from human blastocysts. N Engl J Med 350:1353–1356 (2004).
- de Waard H, de Wit J, Gorgels TG, van den Aardweg G, Andressoo JO, et al: Cell type-specific hypersensitivity to oxidative damage in CSB and XPA mice. DNA Repair 2:13–25 (2003).
- Dofuku R, Biedler JL, Spengler BA, Old LJ: Trisomy of chromosome 15 in spontaneous leukemia of AKR mice. Proc Natl Acad Sci USA 72:1515–1517 (1975).
- Draper JS, Smith K, Gokhale P, Moore HD, Maltby E, et al: Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol 22:53–54 (2004).
- Evans EP, Lyon MF, Daglish M: A mouse translocation giving a metacentric marker chromosome. Cytogenetics 6:105–119 (1967).
- Evans MJ, Kaufman MH: Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156 (1981).
- Forsyth NR, Musio A, Vezzoni P, Simpson AH, Noble BS, McWhir J: Physiologic oxygen enhances human embryonic stem cell clonal recovery and reduces chromosomal abnormalities. Cloning Stem Cells 8:16–23 (2006).
- Hoffman LM, Carpenter MK: Human embryonic stem cell stability. Stem Cell Rev 1:139–144 (2005).
- Hong Y, Stambrook PJ: Restoration of an absent G1 arrest and protection from apoptosis in embryonic stem cells after ionizing radiation. Proc Natl Acad Sci USA 101:14443–14448 (2004).
- Labosky PA, Barlow DP, Hogan BL: Embryonic germ cell lines and their derivation from mouse primordial germ cells. Ciba Found Symp 182:157–168 (1994).
- Liu X, Wu H, Loring J, Hormuzdi S, Disteche CM, et al: Trisomy eight in ES cells is a common potential problem in gene targeting and interferes with germ line transmission. Dev Dyn 209:85–91 (1997).
- Livingstone LR, White A, Sprouse J, Livanos E, Jacks T, Tlsty TD: Altered cell cycle arrest and gene amplification potential accompany loss of wild-type p53. Cell 70:923–935 (1992).
- Longo L, Bygrave A, Grosveld FG, Pandolfi PP: The chromosome make-up of mouse embryonic stem cells is predictive of somatic and germ cell chimaerism. Transgenic Res 6:321–328 (1997).
- Loo DT, Fuquay JI, Rawson CL, Barnes DW: Extended culture of mouse embryo cells without senescence: inhibition by serum. Science 236:200–202 (1987).
- Maitra A, Arking DE, Shivapurkar N, Ikeda M, Stastny V, et al: Genomic alterations in cultured human embryonic stem cells. Nat Genet 37:1099–1103 (2005).
- Martin GR: Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA 78:7634–7638 (1981).
- Mitalipova MM, Rao RR, Hoyer DM, Johnson JA, Meisner LF, et al: Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol 23:19–20 (2005).
- Nagy A, Rossant J, Nagy R, Abramow-Newerly W, Roder JC: Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc Natl Acad Sci USA 90:8424–8428 (1993).
- Neri T, Monti M, Rebuzzini P, Merico V, Garagna S, et al: Mouse fibroblasts are reprogrammed to Oct-4 and Rex-1 gene expression and alkaline phosphatase activity by embryonic stem cell extracts. Cloning Stem Cells 9:394–406 (2007).
- Nichols J, Evans EP, Smith AG: Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity. Development 110:1341–1348 (1990).
- Rice GC, Hoy C, Schimke RT: Transient hypoxia enhances the frequency of dihydrofolate reductase gene amplification in Chinese hamster ovary cells. Proc Natl Acad Sci USA 83:5978–5982 (1986).
- Robertson E, Bradley A, Kuehn M, Evans M: Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature 323:445–448 (1986).
- Savatier P, Huang S, Szekely L, Wiman KG, Samarut J: Contrasting patterns of retinoblastoma protein expression in mouse embryonic stem cells and embryonic fibroblasts. Oncogene 9:809–818 (1994).
- Savatier P, Lapillonne H, Jirmanova L, Vitelli L, Samarut J: Analysis of the cell cycle in mouse embryonic stem cells. Methods Mol Biol 185:27–33 (2002).
- Schnedl W, Dann O, Schweizer D: Effects of counterstaining with DNA binding drugs on fluorescent banding patterns of human and mammalian chromosomes. Eur J Cell Biol 20:290–296 (1980).
- Segev H, Fishman B, Ziskind A, Shulman M, Itskovitz-Eldor J: Differentiation of human embryonic stem cells into insulin-producing clusters. Stem Cells 22:265–274 (2004).
- Stambrook PJ: An ageing question: do embryonic stem cells protect their genomes? Mech Ageing Dev 128:31–35 (2007).
- 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 56:31–34 (2006).
- Sugiyama T, Kurita Y, Nishizuka Y: Chromosome abnormality in rat leukemia induced by 7,12- dimethylbenz[a]anthracene. Science 158:1058–1059 (1967).
- Sumner AT: A simple technique for demonstrating centromeric heterochromatin. Exp Cell Res 75:304–306 (1972).
- Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, et al: Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147 (1998).
- Todaro GJ, Green H: Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol 17:299–313 (1963).
- Van Sloun PP, Jansen JG, Weeda G, Mullenders LH, van Zeeland AA, et al: The role of nucleotide excision repair in protecting embryonic stem cells from genotoxic effects of UV-induced DNA damage. Nucleic Acids Res 27:3276–3282 (1999).
- Wang F, Thirumangalathu S, Loeken MR: Establishment of new mouse embryonic stem cell lines is improved by physiological glucose and oxygen. Cloning Stem Cells 8:108–116 (2006).
- Yin Y, Tainsky MA, Bischoff FZ, Strong LC, Wahl GM: Wild-type p53 restores cell cycle control and inhibits gene amplification in cells with mutant p53 alleles. Cell 70:937–948 (1992).
- Yuasa S, Itabashi Y, Koshimizu U, Tanaka T, Sugimura K, et al: Transient inhibition of BMP signaling by Noggin induces cardiomyocyte differentiation of mouse embryonic stem cells. Nat Biotechnol 23:607–611 (2005).
Article / Publication Details
Copyright / Drug Dosage / DisclaimerCopyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.