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Multiple analysis of mitochondrial metabolism, autophagy and cell death
1Université Paris-Saclay, Institut Gustave Roussy, CNRS, Aspects métaboliques et systémiques de l'oncogénèse pour de nouvelles approches thérapeutiques, Villejuif, France.
Abstract:
In the perspective to evaluate the toxicity of drug candidates or the exploration of intracellular signaling pathways of cell stress response and pathophysiological conditions, we propose to evaluate cell death, autophagy, mitochondrial network and energetic metabolism by a series of optimized joint protocols for neonatal primary rat cardiomyocytes or H9c2 cardiac cell line in 96 well microtiter plates. We used Digitoxigenin and Digoxin, two cardiac glycosides, and Rapamycin as control drugs, for inhibition of oxidative stress-induced cell death and autophagy induction, respectively.
Insights
This study introduces optimized protocols to assess drug toxicity and cell stress responses in cardiac cells. The methods evaluate cell death, autophagy, and metabolism, aiding in drug development and understanding cardiac pathophysiology.
Area of Science:
- Cardiology
- Cell Biology
- Pharmacology
Background:
- Evaluating drug toxicity and cellular stress responses is crucial for understanding cardiac pathophysiology.
- Existing methods may not comprehensively assess multiple cellular parameters simultaneously.
Purpose of the Study:
- To develop and optimize joint protocols for evaluating cell death, autophagy, mitochondrial network, and energetic metabolism in cardiac cells.
- To assess the utility of these protocols in the context of drug toxicity and cell stress response studies.
Main Methods:
- Utilized neonatal primary rat cardiomyocytes and H9c2 cardiac cell lines.
- Developed optimized joint protocols for assessing cell death, autophagy, mitochondrial function, and energy metabolism.
- Employed 96-well microtiter plates for high-throughput analysis.
- Used Digitoxigenin, Digoxin (cardiac glycosides), and Rapamycin as control agents.
Main Results:
- Established a series of optimized protocols for comprehensive analysis of cardiac cell responses.
- Demonstrated the application of these protocols using known modulators of cell death and autophagy.
- Validated the methods for evaluating drug effects on cellular metabolism and mitochondrial networks.
Conclusions:
- The developed protocols provide a robust platform for evaluating drug candidate toxicity and exploring intracellular signaling in cardiac cells.
- These methods facilitate a deeper understanding of cell stress responses and related pathophysiological conditions.
- The approach allows for simultaneous assessment of multiple critical cellular parameters, enhancing research efficiency.
