In-Depth Proteome Analysis Highlights HepaRG Cells as a Versatile Cell System Surrogate for Primary Human Hepatocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. Cell Cultures
2.3. Protein Content and Cell Size of HepaRG, PHH and HepG2
2.4. Karyotyping of HepaRG and HepG2 Cells
2.5. Immunocytochemistry Performed on HepaRG Cells
2.6. Mass Spectrometry Analyses Performed on the HepaRG Cell System
2.6.1. Protein Extraction
2.6.2. Sample Preparation for Intracellular Proteome Analysis
2.6.3. Sample Preparation for Secretome Analysis
2.6.4. Mass Spectrometry Analysis of the Intracellular Proteome
2.6.5. Mass Spectrometry Analysis of the Secretome
2.6.6. Mass Spectrometry Data Processing
2.6.7. Mass Spectrometry Data Analysis
2.6.8. Comparison of the Intracellular Proteome of the Three Cell Systems
2.6.9. Batch-to-Batch Stability of the Proteome of Differentiated HepaRG Cells (HPR116)
3. Results and Discussion
3.1. Main Characteristics of Highly Differentiated HepaRG Cells (HPR116)
3.2. Large-Scale Proteomics of Highly Differentiated HepaRG
3.3. Structural Hotspots of Highly Differentiated HepaRG
3.4. Comprehensive Detoxification Function of Highly Differentiated HepaRG Cells
3.5. Energy Metabolic Functions of Highly Differentiated HepaRG
3.6. Disease-Related Pathways in Highly Differentiated HepaRG, a Promising Field of Application for the HepaRG Cell System
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Tascher, G.; Burban, A.; Camus, S.; Plumel, M.; Chanon, S.; Le Guevel, R.; Shevchenko, V.; Van Dorsselaer, A.; Lefai, E.; Guguen-Guillouzo, C.; et al. In-Depth Proteome Analysis Highlights HepaRG Cells as a Versatile Cell System Surrogate for Primary Human Hepatocytes. Cells 2019, 8, 192. https://doi.org/10.3390/cells8020192
Tascher G, Burban A, Camus S, Plumel M, Chanon S, Le Guevel R, Shevchenko V, Van Dorsselaer A, Lefai E, Guguen-Guillouzo C, et al. In-Depth Proteome Analysis Highlights HepaRG Cells as a Versatile Cell System Surrogate for Primary Human Hepatocytes. Cells. 2019; 8(2):192. https://doi.org/10.3390/cells8020192
Chicago/Turabian StyleTascher, Georg, Audrey Burban, Sandrine Camus, Marine Plumel, Stéphanie Chanon, Remy Le Guevel, Valery Shevchenko, Alain Van Dorsselaer, Etienne Lefai, Christiane Guguen-Guillouzo, and et al. 2019. "In-Depth Proteome Analysis Highlights HepaRG Cells as a Versatile Cell System Surrogate for Primary Human Hepatocytes" Cells 8, no. 2: 192. https://doi.org/10.3390/cells8020192