Cellular Senescence and Aging

1st Edition, Volume 181 - January 1, 2024
Editors: Giulia Petroni, Oliver Kepp, Lorenzo Galluzzi
Language: English
Hardback ISBN:
9 7 8 - 0 - 4 4 3 - 1 8 8 9 6 - 1
eBook ISBN:
9 7 8 - 0 - 4 4 3 - 1 8 8 9 7 - 8

Cellular Senescence and Aging, Volume 181 in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters on t… Read more

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Cellular Senescence and Aging, Volume 181 in the Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters on topics such as assessing polyglutamine tract aggregation in the nematode Caenorhabditis elegans, Generation of glial cell-derived neurotrophic factor (gdnf) morphants in zebrafish larvae by cerebroventricular microinjection of vivo morpholino, Methods for detection of mitochondrial reactive oxygen species in senescent cells, Assessment of cell cycle progression and mitotic slippage by videomicroscopy, The original colorimetric method to detect cellular senescence, and more.

Additional sections cover Assessing microbiota composition in the context of aging, Assessing chronological aging in Saccharomyces cerevisiae, Image processing and supervised machine learning for retinal microglia characterization in senescence, Measuring telomerase activity using TRAP assays, High throughput assessment of cellular senescence, Detection of radiation-induced senescence by the Debacq-Chainiaux protocol: Improvements and upgrade in the detection of positive events, Dynamic and scalable assessment of the senescence-associated secretory phenotype (SASP), Flow cytometry-assisted quantification of cell cycle arrest in cancer cells treated with CDK4/6 inhibitors, and more.

Cover image
Title page
Table of Contents
Series Page
Copyright
Contributors
Cellular senescence and aging at the crossroad between immunity and cancer
Acknowledgments
Conflicts of interest
References
Chapter 1 Assessing polyglutamine tract aggregation in the nematode Caenorhabditis elegans
Abstract
1 Introduction
2 Materials
3 Methods
4 Concluding remarks
5 Notes
Acknowledgments
Author contributions
Conflicts of interest
References
Chapter 2 Generation of glial cell-derived neurotrophic factor (gdnf) morphants in zebrafish larvae by cerebroventricular microinjection of vivo morpholino
Abstract
1 Introduction
2 Materials
3 Optimizing cerebroventricular microinjection of zebrafish larvae
4 Cryosectioning and DAPI counterstaining
5 Cerebroventricular microinjection for knockdown
6 Concluding remarks
7 Notes
References
Chapter 3 Methods for detection of mitochondrial reactive oxygen species in senescent cells
Abstract
1 Introduction
2 Methods for ROS detection in live cells
3 Materials and methods
4 Results and concluding remarks
Acknowledgment
References
Chapter 4 Assessment of cell cycle progression and mitotic slippage by videomicroscopy
Abstract
1 Introduction
2 Materials
3 Methods
4 Notes
5 Concluding remarks
Acknowledgments
Disclosures
References
Chapter 5 The original colorimetric method to detect cellular senescence
Abstract
1 Introduction
2 Materials
3 Methods
4 SA-β-Gal staining solution
5 EdU staining solution
6 SA-β-Gal staining of cells
7 SA-β-Gal staining of tissue samples
8 EdU co-staining
9 Example
10 Concluding remarks
11 Notes
Acknowledgments
References
Chapter 6 Assessing microbiota composition in the context of aging
Abstract
1 Introduction
2 Materials
3 Methods
4 Notes
5 Concluding remarks
Acknowledgments
References
Chapter 7 Assessing chronological aging in Saccharomyces cerevisiae
Abstract
1 Introduction
2 General aspects
3 CLS determination
4 Cell death characterization
5 High-throughput screening
6 Notes
7 Conclusion
References
Chapter 8 Image processing and supervised machine learning for retinal microglia characterization in senescence
Abstract
1 Introduction
2 Retinal dissection
3 Immunostaining and mounting
4 Imaging and image processing
5 SML SVM development for microglia characterization
6 Perivascular microglia quantification
7 Data analysis
8 Concluding remarks
Author Contributions
Acknowledgments
References
Chapter 9 Measuring telomerase activity using TRAP assays
Abstract
1 Telomerase and its effect onto telomeres
2 Telomerase activity in somatic, stem-and cancer cells
3 TRAP as a method for measuring telomerase activity
4 Other non-gel-based methods
5 Quantification of TA and interpretation of results from conventional TRAPs
6 Novel TRAP methods using isothermal amplification and CRISPR-Cas technology
7 Conclusions
References
Chapter 10 High-throughput assessment of cellular senescence
Abstract
1 Introduction
2 Material
3 Methods
4 Concluding remarks
5 Notes
Acknowledgments
Conflicts of interest
Data availability statement
References
Chapter 11 Detection of radiation-induced senescence by the Debacq-Chainiaux protocol: Improvements and upgrade in the detection of positive events
Abstract
1 Introduction
2 Equipment
3 Reagents
4 Irradiation on LINAC
5 HUVECs cell culture
6 Optimization of instrument settings, gating strategy and data acquisition
7 Evaluation of senescence (C12FDG) using spectral flow cytometry
8 Data analysis
9 Overview of the protocol and general considerations
10 Notes
Acknowledgments
Author contributions
Competing interests
References
Chapter 12 Dynamic and scalable assessment of the senescence-associated secretory phenotype (SASP)
Abstract
1 Introduction
2 Materials
3 Methods
4 Notes
Acknowledgments
References
Chapter 13 Flow cytometry-assisted quantification of cell cycle arrest in cancer cells treated with CDK4/6 inhibitors
Abstract
1 Introduction
2 Materials
3 Methods
4 Notes
5. Concluding remarks
Acknowledgment
Competing interests
References
Chapter 14 Mechanistic exploration of autophagy and aging by RNA interference
Abstract
1 Introduction
2 Materials
3 Concluding remarks
4 Notes
Acknowledgments
Conflicts of interest
Data availability statement
References

Giulia Petroni

Giulia Petroni is a PhD Fellow at the Department of Experimental and Clinical Medicine, University of Florence. Giulia does research in Oncology, focusing on the ion channels' role in Colorectal Cancer. She has also an expertise in Clinical Immunology and Immunogenicity. She has published over 15 articles in international peer-reviewed journals.

Affiliations and expertise

Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy

Oliver Kepp

Dr. Oliver Kepp graduated from Humboldt University Berlin and obtained a PhD degree in Molecular Biology. Today he is a tenured researcher at INSERM and a Co-director of the BioCell automation platform at GRCC. Oliver and his team investigate several aspects of tumor immunogenicity, focusing on systems cell biology approaches. He is also a co-founder of Samsara therapeutics and was appointed associate director of the European academy of tumor immunology in 2019. Oliver received the prestigious Clarivate Highly Cited Researcher Award four years in a row since 2018 and has recently been ranked among the top biology and biochemistry scientists in France (#5; www.research.com)

Affiliations and expertise

Equipe labellisée par la Ligue contre le cancer, Université de Paris, Sorbonne Université

Lorenzo Galluzzi

Lorenzo Galluzzi is Assistant Professor of Cell Biology in Radiation Oncology at the Department of Radiation Oncology of the Weill Cornell Medical College, Honorary Assistant Professor Adjunct with the Department of Dermatology of the Yale School of Medicine, Honorary Associate Professor with the Faculty of Medicine of the University of Paris, and Faculty Member with the Graduate School of Biomedical Sciences and Biotechnology of the University of Ferrara, the Graduate School of Pharmacological Sciences of the University of Padova, and the Graduate School of Network Oncology and Precision Medicine of the University of Rome “La Sapienza”. Moreover, he is Associate Director of the European Academy for Tumor Immunology and Founding Member of the European Research Institute for Integrated Cellular Pathology. Galluzzi is best known for major experimental and conceptual contributions to the fields of cell death, autophagy, tumor metabolism and tumor immunology. He has published over 450 articles in international peer-reviewed journals and is the Editor-in-Chief of four journals: OncoImmunology (which he co-founded in 2011), International Review of Cell and Molecular Biology, Methods in Cell biology, and Molecular and Cellular Oncology (which he co-founded in 2013). Additionally, he serves as Founding Editor for Microbial Cell and Cell Stress, and Associate Editor for Cell Death and Disease, Pharmacological Research and iScience.

Affiliations and expertise

Assistant Professor of Cell Biology in Radiation Oncology, Department of Radiation Oncology, Weill Cornell Medical College, NY, USA

Life Sciences

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Cellular Senescence and Aging

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Giulia Petroni

Oliver Kepp

Lorenzo Galluzzi