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Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-Treated SiHa Cervical Cancer Cells
Published on: August 26, 2021
Apoptosis in CHO cell batch cultures: examination by flow cytometry
A Moore1, C J Donahue, J Hooley
1Department of Cell Biology, Genentech. Inc., 94080, South San Francisco, CA, USA.
Cytotechnology
|February 24, 2012
Summary
Chinese hamster ovary cells undergoing apoptosis during stationary phase show significant viability loss. This programmed cell death, primarily occurring in G1/G0 phase, affects up to 60% of cells by culture termination.
Area of Science:
- Biotechnology
- Cell Biology
- Bioprocess Engineering
Background:
- Chinese hamster ovary (CHO) cells are crucial for biopharmaceutical production.
- Maintaining cell viability in batch cultures is essential for optimal protein yield.
- CHO cells in stationary phase exhibit significant viability loss.
Purpose of the Study:
- To investigate the mechanism of cell death in CHO cells during stationary phase.
- To quantify the extent and cell cycle phase of apoptosis.
- To determine if protein secretion affects apoptosis development.
Main Methods:
- Morphological analysis of cell death.
- Flow cytometry with DNA end-labeling (FITC) and propidium iodide staining for cell cycle analysis.
- Radioactive end-labeling of extracted DNA.
Main Results:
- Cell death was predominantly via apoptosis.
- Apoptosis increased significantly in stationary phase, reaching 50-60% by culture end.
- Apoptosis occurred mainly in the G(1)/G(0) phase.
- High DNA FITC end-labeling and hypodiploid propidium iodide binding characterized apoptotic cells.
- Protein secretion did not influence apoptosis progression.
Conclusions:
- Apoptosis is the primary mode of cell death in CHO cells entering stationary phase.
- The G(1)/G(0) phase is a critical window for apoptosis development in these cultures.
- Understanding and mitigating apoptosis is key for improving CHO cell culture productivity.
