skip to Main Content

Induction and Validation of Cellular Senescence in Primary Human Cells.


Maximus Peto’s Commentary

This Journal of Visualized Experiments (JoVE) article looks like a potentially interesting methods paper for working with senescent cells, which might be of interest to some researchers on this distribution list.


Induction and Validation of Cellular Senescence in Primary Human Cells.
J Vis Exp. 2018 Jun 20;(136).
Hernandez-Segura A, Brandenburg S, Demaria M
DOI: 10.3791/57782
PubMed publication date (edat): 7/10/2018

Abstract

Cellular senescence is a state of permanent cell cycle arrest activated in response to different damaging stimuli. Activation of cellular senescence is a hallmark of various pathophysiological conditions including tumor suppression, tissue remodeling and aging. The inducers of cellular senescence in vivo are still poorly characterized. However, a number of stimuli can be used to promote cellular senescence ex vivo. Among them, most common senescence-inducers are replicative exhaustion, ionizing and non-ionizing radiation, genotoxic drugs, oxidative stress, and demethylating and acetylating agents. Here, we will provide detailed instructions on how to use these stimuli to induce fibroblasts into senescence. This protocol can easily be adapted for different types of primary cells and cell lines, including cancer cells. We also describe different methods for the validation of senescence induction. In particular, we focus on measuring the activity of the lysosomal enzyme Senescence-Associated β-galactosidase (SA-β-gal), the rate of DNA synthesis using 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay, the levels of expression of the cell cycle inhibitors p16 and p21, and the expression and secretion of members of the Senescence-Associated Secretory Phenotype (SASP). Finally, we provide example results and discuss further applications of these protocols.

PMID: 29985363
Free Full-Text:

Maximus Peto

Max Peto is a longevity researcher and founder of Long Life Labs. A biochemist by training, he studies the biochemistry of aging and longevity and has worked with research organizations such as SENS Research Foundation, Methuselah Foundation, BioAge Labs, Life Extension Foundation, and Ichor Therapeutics. His work at Long Life Labs is focused on empowering people to understand and manage the most critical factors for better health and longer life.

Free report on high blood pressure

The American Heart Association estimates more than 100 million Americans have high blood pressure, also known as “hypertension”. Learn more about the cause of high blood pressure and how you can reverse it in our free report.

Coming soon. We’ll send you the report as soon as it’s published.

*We do not share your email address with anyone.

Free longevity biomarker report

Biomarker levels predict the risk of early death—and we can change them! Learn about some important longevity biomarkers in our free report.

Coming soon. We’ll send you the report as soon as it’s published.

*We do not share your email address with anyone.

Free diabetes report

An estimated 50% of American adults have either prediabetes or type 2 diabetes. Learn more about the cause of type 2 diabetes, prediabetes, insulin resistance, and how to reverse them in our free report.

Coming soon. We’ll send you the report as soon as it’s published.

*We do not share your email address with anyone.

Back To Top