Embryonic stem (ES) cells are pluripotent cells widely used in cell therapy and tissue engineering. However, the broader clinical applications of ES cells are limited by their genomic instability and karyotypic abnormalities. Thus, understanding the mechanisms underlying ES cell karyotypic abnormalities is critical to optimizing their clinical use. In this study, we focused on proliferating human and mouse ES cells undergoing multipolar divisions. Specifically, we analyzed the frequency and outcomes of such divisions using a combination of time-lapse microscopy and cell tracking. This revealed that cells resulting from multipolar divisions were not only viable, but they also frequently underwent subsequent cell divisions. Our novel data also showed that in human and mouse ES cells, multipolar spindles allowed more robust escape from chromosome segregation control mechanisms than bipolar spindles. Considering the frequency of multipolar divisions in proliferating ES cells, it is conceivable that cell division errors underlie ES cell karyotypic instability.

Institute of Biophysics of the Czech Academy of Sciences Brno Czech Republic

Masaryk University CEITEC Central European Institute of Technology Cellular Imaging Core Facility Brno Czech Republic

Masaryk University Faculty of Medicine Department of Histology and Embryology Brno Czech Republic

Pasteur Institute Image Analysis Hub Paris France

St Anne's University Hospital International Clinical Research Center Brno Czech Republic

Zobrazit více v PubMed

Ballabeni A, Park IH, Zhao R, Wang W, Lerou PH, Daley GQ, Kirschner MW (2011). Cell cycle adaptations of embryonic stem cells. Proc Natl Acad Sci U S A 108(48): 19252-19257. DOI: 10.1073/pnas.1116794108. PubMed DOI

Brinkley BR (2001). Managing the centrosome numbers game: from chaos to stability in cancer cell division. Trends Cell Biol 11(1): 18-21. DOI: 10.1016/s0962-8924(00)01872-9. PubMed DOI

Castedo M, Perfettini JL, Roumier T, Valent A, Raslova H, Yakushijin K, et al. (2004). Mitotic catastrophe constitutes a special case of apoptosis whose suppression entails aneuploidy. Oncogene 23(25): 4362-4370. DOI: 10.1038/sj.onc.1207572. PubMed DOI

Celton-Morizur S, Merlen G, Couton D, Margall-Ducos G, Desdouets C (2009). The insulin/Akt pathway controls a specific cell division program that leads to generation of binucleated tetraploid liver cells in rodents. J Clin Invest 119(7): 1880-1887. DOI: 10.1172/jci38677. PubMed DOI

Chen J, Liu J (2015). Erroneous silencing of the mitotic checkpoint by aberrant spindle pole-kinetochore coordination. Biophys J 109(11): 2418-2435. DOI: 10.1016/j.bpj.2015.10.024. PubMed DOI

Draper JS, Smith K, Gokhale P, Moore HD, Maltby E, Johnson J, et al. (2004). Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nat Biotechnol 22(1): 53-54. DOI: 10.1038/nbt922. PubMed DOI

Duncan AW, Taylor MH, Hickey RD, Hanlon Newell AE, Lenzi ML, Olson SB, et al. (2010). The ploidy conveyor of mature hepatocytes as a source of genetic variation. Nature 467(7316): 707-710. DOI: 10.1038/nature09414. PubMed DOI

Durrbaum M, Storchova Z (2016). Effects of aneuploidy on gene expression: implications for cancer. FEBS J 283(5): 791-802. DOI: 10.1111/febs.13591. PubMed DOI

Foley EA, Kapoor TM (2013). Microtubule attachment and spindle assembly checkpoint signalling at the kinetochore. Nat Rev Mol Cell Biol 14(1): 25-37. DOI: 10.1038/nrm3494. PubMed DOI

Galimberti F, Thompson SL, Ravi S, Compton DA, Dmitrovsky E (2011). Anaphase catastrophe is a target for cancer therapy. Clin Cancer Res 17(6): 1218-1222. DOI: 10.1158/1078-0432.CCR-10-1178. PubMed DOI

Ganem NJ, Cornils H, Chiu SY, O'Rourke KP, Arnaud J, Yimlamai D, et al. (2014). Cytokinesis failure triggers hippo tumor suppressor pathway activation. Cell 158(4): 833-848. DOI: 10.1016/j.cell.2014.06.029. PubMed DOI

Ganem NJ, Godinho SA, Pellman D (2009). A mechanism linking extra centrosomes to chromosomal instability. Nature 460(7252): 278-282. DOI: 10.1038/nature08136. PubMed DOI

Gentric G, Desdouets C (2014). Polyploidization in liver tissue. Am J Pathol 184(2): 322-331. DOI: 10.1016/j.ajpath.2013.06.035. PubMed DOI

Gisselsson D, Jin Y, Lindgren D, Persson J, Gisselsson L, Hanks S, et al. (2010). Generation of trisomies in cancer cells by multipolar mitosis and incomplete cytokinesis. Proc Natl Acad Sci U S A 107(47): 20489-20493. DOI: 10.1073/pnas.1006829107. PubMed DOI

Holubcova Z, Blayney M, Elder K, Schuh M (2015). Human oocytes. Error-prone chromosome-mediated spindle assembly favors chromosome segregation defects in human oocytes. Science 348(6239): 1143-1147. DOI: 10.1126/science.aaa9529. PubMed DOI

Holubcova Z, Matula P, Sedlackova M, Vinarsky V, Dolezalova D, Barta T, et al. (2011). Human embryonic stem cells suffer from centrosomal amplification. Stem Cells 29(1): 46-56. DOI: 10.1002/stem.549. PubMed DOI

Imreh MP, Gertow K, Cedervall J, Unger C, Holmberg K, Szoke K, et al. (2006). In vitro culture conditions favoring selection of chromosomal abnormalities in human ES cells. J Cell Biochem 99(2): 508-516. DOI: 10.1002/jcb.20897. PubMed DOI

International Stem Cell Initiative, Amps K, Andrews PW, Anyfantis G, Armstrong L, Avery S, et al. (2011). Screening ethnically diverse human embryonic stem cells identifies a chromosome 20 minimal amplicon conferring growth advantage. Nat Biotechnol 29(12): 1132-1144. DOI: 10.1038/nbt.2051. PubMed DOI

Kallas A, Pook M, Maimets M, Zimmermann K, Maimets T (2011). Nocodazole treatment decreases expression of pluripotency markers Nanog and Oct4 in human embryonic stem cells. PloS One 6(4): e19114. DOI: 10.1371/journal.pone.0019114. PubMed DOI

Kwon M, Godinho SA, Chandhok NS, Ganem NJ, Azioune A, Thery M, Pellman D (2008). Mechanisms to suppress multipolar divisions in cancer cells with extra centrosomes. Genes Dev 22(16): 2189-2203. DOI: 10.1101/gad.1700908. PubMed DOI

Lin TX, Chao C, Saito S, Mazur SJ, Murphy ME, Appella E, Xu Y (2005). P53 induces differentiation of mouse embryonic stem cells by suppressing Nanog expression. Nature Cell Biol 7(2): 165-171. DOI: 10.1038/ncb1211. PubMed DOI

Martello G, Smith A (2014). The nature of embryonic stem cells. Annu Rev Cell Dev Biol 30: 647-675. DOI: 10.1146/annurev-cellbio-100913-013116. PubMed DOI

Nakagawa S, FitzHarris G (2017). Intrinsically defective microtubule dynamics contribute to age-related chromosome segregation errors in mouse oocyte meiosis-I. Curr Biol 27(7): 1040-1047. DOI: 10.1016/j.cub.2017.02.025. PubMed DOI

Park Y, Depeursinge C, Popescu G (2018). Quantitative phase imaging in biomedicine. Nature Photonics 12(10): 578-589. DOI: 10.1038/s41566-018-0253-x. DOI

Poser I, Sarov M, Hutchins JR, Heriche JK, Toyoda Y, Pozniakovsky A, et al. (2008). BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals. Nat Methods 5(5): 409-415. DOI: 10.1038/nmeth.1199. PubMed DOI

Quintyne NJ, Reing JE, Hoffelder DR, Gollin SM, Saunders WS (2005). Spindle multipolarity is prevented by centrosomal clustering. Science 307(5706): 127-129. DOI: 10.1126/science.1104905. PubMed DOI

Ring D, Hubble R, Kirschner M (1982). Mitosis in a cell with multiple centrioles. J Cell Biol 94(3): 549-556. DOI: 10.1083/jcb.94.3.549. PubMed DOI

Sanchez-Aguilera A, Montalban C, de la Cueva P, Sanchez-Verde L, Morente MM, Garcia-Cosio M, et al. (2006). Tumor microenvironment and mitotic checkpoint are key factors in the outcome of classic Hodgkin lymphoma. Blood 108(2): 662-668. DOI: 10.1182/blood-2005-12-5125. PubMed DOI

Santaguida S, Richardson A, Iyer DR, M'Saad O, Zasadil L, Knouse KA, et al. (2017). Chromosome mis-segregation generates cell-cycle-arrested cells with complex karyotypes that are eliminated by the immune system. Dev Cell 41(6): 638-651.e5. DOI: 10.1016/j.devcel.2017.05.022. PubMed DOI

Silkworth WT, Cimini D (2012). Transient defects of mitotic spindle geometry and chromosome segregation errors. Cell Div 7(1): 19. DOI: 10.1186/1747-1028-7-19. PubMed DOI

Silkworth WT, Nardi IK, Scholl LM, Cimini D (2009). Multipolar spindle pole coalescence is a major source of kinetochore mis-attachment and chromosome mis-segregation in cancer cells. Plos One 4(8): e6564. DOI: 10.1371/journal.pone.0006564. PubMed DOI

Sluder G, Thompson EA, Miller FJ, Hayes J, Rieder CL (1997). The checkpoint control for anaphase onset does not monitor excess numbers of spindle poles or bipolar spindle symmetry. J Cell Sci 110(Pt 4): 421-429. PubMed

Spits C, Mateizel I, Geens M, Mertzanidou A, Staessen C, Vandeskelde Y, et al. (2008). Recurrent chromosomal abnormalities in human embryonic stem cells. Nat Biotechnol 26(12): 1361-1363. DOI: 10.1038/nbt.1510. PubMed DOI

Storchova Z, Kuffer C (2008). The consequences of tetraploidy and aneuploidy. J Cell Sci 121(Pt 23): 3859-3866. DOI: 10.1242/jcs.039537. PubMed DOI

Vitale I, Galluzzi L, Castedo M, Kroemer G (2011). Mitotic catastrophe: a mechanism for avoiding genomic instability. Nat Rev Mol Cell Biol 12(6): 385-392. DOI: 10.1038/nrm3115. PubMed DOI

Werbowetski-Ogilvie TE, Bosse M, Stewart M, Schnerch A, Ramos-Mejia V, Rouleau A, et al. (2009). Characterization of human embryonic stem cells with features of neoplastic progression. Nat Biotechnol 27(1): 91-97. DOI: 10.1038/nbt.1516. PubMed DOI

Yi Q, Zhao X, Huang Y, Ma T, Zhang Y, Hou H, et al. (2011). P53 dependent centrosome clustering prevents multipolar mitosis in tetraploid cells. PLoS One 6(11): e27304. DOI: 10.1371/journal.pone.0027304. PubMed DOI

Zhang M, Cheng L, Jia Y, Liu G, Li C, Song S, et al. (2016). Aneuploid embryonic stem cells exhibit impaired differentiation and increased neoplastic potential. EMBO J 35(21): 2285-2300. DOI: 10.15252/embj.201593103. PubMed DOI

Zhang ZN, Chung SK, Xu Z, Xu Y (2014). Oct4 maintains the pluripotency of human embryonic stem cells by inactivating p53 through Sirt1-mediated deacetylation. Stem Cells 32(1): 157-165. DOI: 10.1002/stem.1532. PubMed DOI

Geometric Control of Cell Behavior by Biomolecule Nanodistribution