Using cell culture models of centrosome amplification to study centrosome clustering in cancer

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

2 Scopus citations


The link between centrosome amplification and cancer has been recognized for more than a century, raising many key questions about the biology of both normal and cancer cells. In particular, the presence of extra centrosomes imposes a great challenge to a dividing cell by increasing the likelihood of catastrophic multipolar divisions. Only recently have we begun to understand how cancer cells successfully divide by clustering their extra centrosomes for bipolar division. Several hurdles to dissecting centrosome clustering include limitations in the methodologies used to quantify centrosome amplification, and the lack of appropriate cell culture models. Here, we describe how to accurately assess centrosome number and create isogenic cell lines with or without centrosome amplification. We then describe how imaging of cell division in these cell culture models leads to identification of the molecular machinery uniquely required for cells with extra centrosomes. These approaches have led to the identification of molecular targets for selective cancer therapeutics that can kill cancer cells with extra centrosomes without affecting normal cells with two centrosomes. We further anticipate that the approaches described here will be broadly applicable for studying the causes and consequences of centrosome amplification in a variety of contexts across cancer pathophysiology, such as cell migration and metastasis.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages26
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745

Bibliographical note

Funding Information:
This protocol is currently used in David Pellman laboratory and has been refined over the years by several people including Drs. S. Godinho and N. Ganem. We thank N. Umbreit, A. Spektor, Y. Kaplan, H. Zhang for discussions and/or comments on the manuscript; all members of the Pellman laboratory for advice and suggestions. M.K was supported by a special fellow award from the Leukemia and Lymphoma Society and a Susan G. Komen grant. Our work was supported by the Howard Hughes Medical Institute and the NIH (GM061345). Mijung Kwon

Publisher Copyright:
© Springer Science+Business Media New York 2016.


  • Aneuploidy
  • Cancer
  • Cell division
  • Centrosome
  • Centrosome amplification
  • Extra centrosomes
  • Mitosis
  • Multipolar spindle
  • Selective cancer therapeutics


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