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Assay for Assessing Mitochondrial Function in iPSC-Derived Neural Stem Cells and Dopaminergic Neurons
Gourav Roy-Choudhury1, Marcel M Daadi2,3
1Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX, USA.
Methods in Molecular Biology (Clifton, N.J.)
|January 19, 2019
Summary
Assessing mitochondrial function in induced pluripotent stem cell-derived neural stem cells using Seahorse technology is crucial for drug discovery. This method enables reliable evaluation of compounds targeting mitochondrial dysfunction.
Area of Science:
- Biochemistry
- Cell Biology
- Neuroscience
Background:
- Mitochondrial dysfunction is implicated in various diseases.
- Induced pluripotent stem cells (iPSCs) offer a patient-specific disease modeling platform.
- Neural stem cells (NSCs) derived from iPSCs (iPSC-NSCs) can recapitulate disease phenotypes.
Purpose of the Study:
- To establish and validate a method for assessing mitochondrial bioenergetics in iPSC-NSCs.
- To utilize the Seahorse XFe96 analyzer for high-throughput drug screening of compounds targeting mitochondrial function.
- To demonstrate the utility of iPSC-NSCs as a cellular model for studying mitochondrial dysfunction.
Main Methods:
- Generation of iPSC-NSCs from patient-derived iPSCs.
- Culture and maintenance of iPSC-NSCs.
- Assessment of mitochondrial respiration and glycolysis using the Seahorse XFe96 analyzer.
Main Results:
- Demonstrated the feasibility of measuring mitochondrial bioenergetics in iPSC-NSCs.
- Established a reliable protocol for Seahorse analysis in this cellular model.
- Showcased the potential for phenotypic screening of drug candidates.
Conclusions:
- Seahorse analysis of iPSC-NSCs provides a robust platform for studying mitochondrial dysfunction.
- This approach facilitates drug discovery for diseases associated with mitochondrial impairment.
- iPSC-NSCs are a valuable tool for personalized medicine and therapeutic development.
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