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Related Experiment Video

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A cGMP-applicable Expansion Method for Aggregates of Human Neural Stem and Progenitor Cells Derived From Pluripotent Stem Cells or Fetal Brain Tissue

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Human embryonic stem cells suffer from centrosomal amplification.

Zuzana Holubcová¹, Pavel Matula, Miroslava Sedláčková

¹Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.

Stem Cells (Dayton, Ohio)

|October 21, 2010

Summary

Cultured human embryonic stem cells (hESCs) often exhibit supernumerary centrosomes, increasing risk for chromosome instability. This condition is linked to undifferentiated states and improves with better cell attachment and regulated cell division factors.

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Area of Science:

Cell Biology
Stem Cell Biology
Genetics

Background:

Human embryonic stem cells (hESCs) propagation in culture can lead to karyotype alterations.
Chromosomal instability in malignancies is often linked to centrosome overamplification.
Centrosome amplification disrupts balanced chromosome segregation.

Purpose of the Study:

To investigate the presence and causes of supernumerary centrosomes in cultured hESCs.
To determine the relationship between centrosome amplification and hESC state.
To identify factors contributing to centrosome overamplification in hESCs.

Main Methods:

Microscopy to observe centrosomes during mitosis in hESCs.
Analysis of centrosome number in relation to hESC differentiation state.
Investigating the effect of cell substratum attachment and specific kinase inhibitors (CDK2, Aurora A) on centrosome amplification.

Main Results:

A high percentage of cultured hESCs displayed supernumerary centrosomes during mitosis.
Supernumerary centrosomes were associated with the undifferentiated hESC state and decreased with prolonged culture.
Improved cell attachment and inhibition of CDK2/Aurora A reduced multicentrosomal mitoses.
Both cell cycle overduplication and mitotic failure contribute to numerical centrosomal abnormalities.

Conclusions:

Supernumerary centrosomes are a significant risk factor for chromosome instability in cultured hESCs.
Attenuated cell attachment and deregulation of centrosome number control contribute to centrosome amplification.
Centrosome abnormalities in hESCs require evaluation for safe clinical applications.