The Use of Plasmid Microinjection to Study Specific Cell Cycle Phase Transitions

1999 ◽  
pp. 31-43
Author(s):  
E. S. Knudsen
Keyword(s):  
Cell Cycle ◽  
Phase Transitions ◽  
Cell Cycle Phase ◽  
Cycle Phase ◽  
Specific Cell

scholarly journals Data Driven Cell Cycle Model to Quantify the Efficacy of Cancer Therapeutics Targeting Specific Cell-Cycle Phases From Flow Cytometry Results

2021 ◽  
Vol 1 ◽  
Author(s):  
David W. James ◽  
Andrew Filby ◽  
M. Rowan Brown ◽  
Huw D. Summers ◽  
Lewis W. Francis ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Phase Transitions ◽  
Mechanism Of Action ◽  
Cell Cycle Phase ◽  
Data Driven ◽  
Cycle Phase ◽  
Cell Populations ◽  
Specific Cell ◽  
Cell Cycle Phases

Many chemotherapeutic drugs target cell processes in specific cell cycle phases. Determining the specific phases targeted is key to understanding drug mechanism of action and efficacy against specific cancer types. Flow cytometry experiments, combined with cell cycle phase and division round specific staining, can be used to quantify the current cell cycle phase and number of mitotic events of each cell within a population. However, quantification of cell interphase times and the efficacy of cytotoxic drugs targeting specific cell cycle phases cannot be determined directly. We present a data driven computational cell population model for interpreting experimental results, where in-silico populations are initialized to match observable results from experimental populations. A two-stage approach is used to determine the efficacy of cytotoxic drugs in blocking cell-cycle phase transitions. In the first stage, our model is fitted to experimental multi-parameter flow cytometry results from untreated cell populations to identify parameters defining probability density functions for phase transitions. In the second stage, we introduce a blocking routine to the model which blocks a percentage of attempted transitions between cell-cycle phases due to therapeutic treatment. The resulting model closely matches the percentage of cells from experiment in each cell-cycle phase and division round. From untreated cell populations, interphase and intermitotic times can be inferred. We then identify the specific cell-cycle phases that cytotoxic compounds target and quantify the percentages of cell transitions that are blocked compared with the untreated population, which will lead to improved understanding of drug efficacy and mechanism of action.

scholarly journals Measurement of cell-cycle phase-specific cell death using Hoechst 33342 and propidium iodide: preservation by ethanol fixation.

1988 ◽  
Vol 36 (9) ◽  
pp. 1147-1152 ◽  
Author(s):  
G Ciancio ◽  
A Pollack ◽  
M A Taupier ◽  
N L Block ◽  
G L Irvin
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Propidium Iodide ◽  
Cell Cycle Phase ◽  
Cytotoxic Agents ◽  
Cycle Phase ◽  
Specific Cell ◽  
Dead Cell ◽  
Hoechst 33342 ◽  
The Dead

We developed a rapid technique for preservation of Hoechst 33342/propidium iodide-stained cells, using ethanol as a fixative. Combined staining with these dyes makes possible analysis of cell-cycle phase-specific cell death. The technique relies on exclusion of propidium iodide from the viable cells, whereas Hoechst stains all of the cells. The bivariate histograms resulting from the flow cytometric analysis contain the equivalent of two single-parameter DNA histograms, one of the living and the other of the dead cell population. Preservation of staining involved addition of 25% ethanol in PBS after propidium iodide staining and before Hoechst staining. The separation between the living and the dead cell populations was maintained for over 3 days at 4 degrees C. This technique will be valuable for quantitative evaluation of the cell-cycle phase-specific effects of cytostatic or cytotoxic agents, particularly in situations where a lag period between staining and analysis is unavoidable.

scholarly journals DNA damage checkpoint dynamics drive cell cycle phase transitions

2017 ◽  
Author(s):  
Hui Xiao Chao ◽  
Cere E. Poovey ◽  
Ashley A. Privette ◽  
Gavin D. Grant ◽  
Hui Yan Chao ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Damage ◽  
Phase Transitions ◽  
Cell Cycle Progression ◽  
S Phase ◽  
Cell Cycle Phase ◽  
Cycle Phase ◽  
Dna Damage Checkpoint ◽  
Cycle Progression ◽  
Dna Damage Checkpoints

ABSTRACTDNA damage checkpoints are cellular mechanisms that protect the integrity of the genome during cell cycle progression. In response to genotoxic stress, these checkpoints halt cell cycle progression until the damage is repaired, allowing cells enough time to recover from damage before resuming normal proliferation. Here, we investigate the temporal dynamics of DNA damage checkpoints in individual proliferating cells by observing cell cycle phase transitions following acute DNA damage. We find that in gap phases (G1 and G2), DNA damage triggers an abrupt halt to cell cycle progression in which the duration of arrest correlates with the severity of damage. However, cells that have already progressed beyond a proposed “commitment point” within a given cell cycle phase readily transition to the next phase, revealing a relaxation of checkpoint stringency during later stages of certain cell cycle phases. In contrast to G1 and G2, cell cycle progression in S phase is significantly less sensitive to DNA damage. Instead of exhibiting a complete halt, we find that increasing DNA damage doses leads to decreased rates of S-phase progression followed by arrest in the subsequent G2. Moreover, these phase-specific differences in DNA damage checkpoint dynamics are associated with corresponding differences in the proportions of irreversibly arrested cells. Thus, the precise timing of DNA damage determines the sensitivity, rate of cell cycle progression, and functional outcomes for damaged cells. These findings should inform our understanding of cell fate decisions after treatment with common cancer therapeutics such as genotoxins or spindle poisons, which often target cells in a specific cell cycle phase.

scholarly journals Selective protein degradation: a rheostat to modulate cell-cycle phase transitions

2013 ◽  
Vol 65 (10) ◽  
pp. 2603-2615 ◽  
Author(s):  
Pascal Genschik ◽  
Katia Marrocco ◽  
Lien Bach ◽  
Sandra Noir ◽  
Marie-Claire Criqui
Keyword(s):  
Cell Cycle ◽  
Phase Transitions ◽  
Protein Degradation ◽  
Cell Cycle Phase ◽  
Cycle Phase

scholarly journals Accurate delineation of cell cycle phase transitions in living cells with PIP-FUCCI

Cell Cycle ◽  
2018 ◽  
Vol 17 (21-22) ◽  
pp. 2496-2516 ◽  
Author(s):  
Gavin D. Grant ◽  
Katarzyna M. Kedziora ◽  
Juanita C. Limas ◽  
Jeanette Gowen Cook ◽  
Jeremy E. Purvis
Keyword(s):  
Cell Cycle ◽  
Phase Transitions ◽  
Living Cells ◽  
Cell Cycle Phase ◽  
Cycle Phase

Identification of age-structured models: Cell cycle phase transitions

2008 ◽  
Vol 99 (4) ◽  
pp. 960-974 ◽  
Author(s):  
E. Sherer ◽  
E. Tocce ◽  
R.E. Hannemann ◽  
A.E. Rundell ◽  
D. Ramkrishna
Keyword(s):  
Cell Cycle ◽  
Phase Transitions ◽  
Cell Cycle Phase ◽  
Cycle Phase ◽  
Age Structured

Growth and cell cycle phase composition of fibroblasts in the reconstituted dermis model.

1990 ◽  
Vol 52 (5) ◽  
pp. 986-992
Author(s):  
Takeshi KONO ◽  
Tsukasa TANII ◽  
Masayoshi FURUKAWA ◽  
Nobuyuki MIZUNO ◽  
Shoji TANIGUCHI ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Phase Composition ◽  
Cell Cycle Phase ◽  
Cycle Phase
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