Human Embryonic Stem Cells Suffer from Centrosomal Amplification§

Authors

  • Zuzana Holubcová,

    1. Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    Search for more papers by this author
  • Pavel Matula,

    1. Centre for Biomedical Image Analysis, Faculty of Informatics, Masaryk University, Brno, Czech Republic
    Search for more papers by this author
  • Miroslava Sedláčková,

    1. Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    Search for more papers by this author
  • Vladimír Vinarský,

    1. Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    2. Department of Molecular Embryology, Institute of Experimental Medicine, v.v.i., Academy of Sciences of the Czech Republic, Brno, Czech Republic
    Search for more papers by this author
  • Dáša Doležalová,

    1. Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    2. Department of Molecular Embryology, Institute of Experimental Medicine, v.v.i., Academy of Sciences of the Czech Republic, Brno, Czech Republic
    Search for more papers by this author
  • Tomáš Bárta,

    1. Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    2. Department of Molecular Embryology, Institute of Experimental Medicine, v.v.i., Academy of Sciences of the Czech Republic, Brno, Czech Republic
    Search for more papers by this author
  • Petr Dvořák,

    1. Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    2. Department of Molecular Embryology, Institute of Experimental Medicine, v.v.i., Academy of Sciences of the Czech Republic, Brno, Czech Republic
    Search for more papers by this author
  • Aleš Hampl

    Corresponding author
    1. Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic
    2. Department of Molecular Embryology, Institute of Experimental Medicine, v.v.i., Academy of Sciences of the Czech Republic, Brno, Czech Republic
    • Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Kamenice 5, Brno 62500, Czech Republic
    Search for more papers by this author
    • Telephone: 420-549-49-3514; Fax: 420-549-49-8001


  • Author contributions: Z.H.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; P.M.: collection and assembly of data, data analysis and interpretation; M.S.: collection and assembly of data, data analysis and interpretation; V.V.: data analysis and interpretation; D.D.: data analysis and interpretation; T.B.: data analysis and interpretation; P.D.: data analysis and interpretation; A.H.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript.

  • First published online in STEM CELLS EXPRESS October 19, 2010.

  • §

    Disclosure of potential conflicts of interest is found at the end of this article.

Abstract

Propagation of human embryonic stem cells (hESCs) in culture tends to alter karyotype, potentially limiting the prospective use of these cells in patients. The chromosomal instability of some malignancies is considered to be driven, at least in part, by centrosomal overamplification, perturbing balanced chromosome segregation. Here, we report, for the first time, that very high percentage of cultured hESCs has supernumerary centrosomes during mitosis. Supernumerary centrosomes were strictly associated with an undifferentiated hESC state and progressively disappeared on prolonged propagation in culture. Improved attachment to culture substratum and inhibition of CDK2 and Aurora A (key regulators of centrosomal metabolism) diminished the frequency of multicentrosomal mitoses. Thus, both attenuated cell attachment and deregulation of machinery controlling centrosome number contribute to centrosomal overamplification in hESCs. Linking the excessive number of centrosomes in mitoses to the ploidy indicated that both overduplication within a single cell cycle and mitotic failure contributed to generation of numerical centrosomal abnormalities in hESCs. Collectively, our data indicate that supernumerary centrosomes are a significant risk factor for chromosome instability in cultured hESCs and should be evaluated when new culture conditions are being implemented. STEM CELLS 2011;29:46–56

Ancillary