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Updated: Jul 4, 2025
Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations
Published on: September 20, 2019
Assessment of cell cycle progression and mitotic slippage by videomicroscopy
Luca Mattiello1, Sara Soliman Abdel Rehim2, Gwenola Manic1
1IIGM-Italian Institute for Genomic Medicine, c/o IRCSS, Candiolo, Italy; Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Italy.
Abstract:
Senescence is a state of irreversible cell cycle arrest accompanied by the acquisition of the senescence-associated secretory phenotype (SASP), which is activated in response to a variety of damaging stimuli, including genotoxic therapy. Accumulating evidence indicates that mitotic stress also promotes entry into senescence. This occurs via a mechanism involving defective mitoses and mitotic arrest, followed by abortion of cell division and slippage in the G1 phase. In this process, mitotic slippage leads to the generation of senescent cells characterized by a large cell body and a multinucleated and/or enlarged nuclear size. Here, we provide a detailed protocol for the assessment of cell proliferation and mitotic slippage in colorectal cancer cells upon pharmacological inhibition of the mitotic kinesin KIF11, best known as EG5. This approach can be used for preliminary characterization of senescence induction by therapeutics, but requires validation with standard senescence assays.
Insights
This study details a method to assess cell proliferation and mitotic slippage in colorectal cancer cells. This protocol aids in characterizing therapeutic induction of senescence, a state of irreversible cell cycle arrest.
Area of Science:
- Cell Biology
- Cancer Research
- Molecular Biology
Background:
- Senescence is a state of irreversible cell cycle arrest linked to the senescence-associated secretory phenotype (SASP).
- Genotoxic therapy and mitotic stress can induce senescence.
- Mitotic slippage, characterized by defective mitosis and cell cycle progression, generates senescent cells with distinct morphological features.
Purpose of the Study:
- To provide a detailed protocol for assessing cell proliferation and mitotic slippage.
- To investigate senescence induction in colorectal cancer cells.
- To evaluate the effect of KIF11 (EG5) inhibition on cell cycle progression and senescence.
Main Methods:
- Pharmacological inhibition of the mitotic kinesin KIF11 (EG5) in colorectal cancer cells.
- Assessment of cell proliferation.
- Evaluation of mitotic slippage, including multinucleation and enlarged nuclear size.
Main Results:
- The protocol allows for the assessment of cell proliferation and mitotic slippage.
- Mitotic slippage leads to characteristic senescent cell morphology.
- This method can be used for preliminary characterization of senescence induction by therapeutics.
Conclusions:
- The developed protocol offers a method to study mitotic slippage and its role in senescence.
- This approach can serve as a preliminary tool for evaluating senescence induction by novel therapeutics.
- Further validation with standard senescence assays is recommended.
