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Pitfalls and requirements in quantifying asymmetric mitotic segregation.
Journal article

Pitfalls and requirements in quantifying asymmetric mitotic segregation.

  • Loeffler D Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule Zurich, Basel, Switzerland.
  • Schneiter F Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule Zurich, Basel, Switzerland.
  • Schroeder T Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule Zurich, Basel, Switzerland.
  • 2019-12-10
Published in:
  • Annals of the New York Academy of Sciences. - 2020
NUMB
asymmetric cell division
asymmetric cell fate
asymmetric inheritance
hematopoietic stem cells
stem cell
Animals
Asymmetric Cell Division
Cell Differentiation
Cell Division
Cells, Cultured
Hematopoietic Stem Cells
Inheritance Patterns
Male
Membrane Proteins
Mice
Mice, Inbred C57BL
Microtubules
Mitosis
Nerve Tissue Proteins
Nocodazole
Polymerization
Tissue Distribution
Tubulin Modulators
English The asymmetric inheritance of NUMB during mitosis determines future daughter cell fates in multiple model organisms. NUMB asymmetric inheritance has also been postulated for hematopoietic stem cell (HSC) divisions but remained controversial until recently. To reconcile conflicting reports, we revisited the evidence for asymmetric inheritance of NUMB during HSC divisions. We demonstrate that previously used strategies to identify dividing cells in fixed samples suffer from multiple systematic errors. Nonmitotic cells in close proximity are frequently mistaken as dividing cells, while mitotic cells are not detected. Furthermore, microtubule depolymerization by either nocodazole or low temperatures prevents the reliable detection of mitosis and introduces mitotic artifacts. Without artificial microtubule depolymerization and by the use of reliable mitotic markers, we find NUMB differences in daughter cells to be reduced and restricted to cells with low NUMB expression and thus low signal over background. This bias fits the expected random distribution of simulated noise data, suggesting that the putative asymmetric inheritance of NUMB in HSCs could be merely technical noise. We conclude that functionally relevant asymmetric inheritance of NUMB and other factors in mitotic HSCs and other cells cannot be conclusively demonstrated using snapshot data and requires alternative approaches, such as continuous quantitative single-cell analysis.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/3789
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