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Related Experiment Video

Updated: Sep 1, 2025

SA-β-Galactosidase-Based Screening Assay for the Identification of Senotherapeutic Drugs
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A protocol for rapid construction of senescent cells.

Xing Yu1,2, Jing Quan1, Shuai Chen3

  • 1Key Laboratory of Molecular Epidemiology of Hunan Province, School of Medicine, Hunan Normal University, Changsha, China.

Frontiers in Integrative Neuroscience
|August 15, 2022
PubMed
Summary

Cellular senescence, a key aging mechanism, drives age-related diseases. This study develops a rapid in vitro model using conditioned medium from senescent hepatocytes to induce senescence, aiding disease mechanism research.

Keywords:
SASPagingbiomarkercellular senescencesenescent cell models

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Area of Science:

  • Gerontology and Cellular Biology
  • Molecular mechanisms of aging and disease

Background:

  • Aging is a major risk factor for chronic diseases, with cellular senescence implicated in age-related conditions.
  • Senescent cells, characterized by cell cycle arrest and a pro-inflammatory secretory phenotype (SASP), accumulate with age.
  • While beneficial roles exist, senescent cell accumulation contributes to tissue dysfunction and age-associated diseases.

Purpose of the Study:

  • To establish a rapid in vitro model for studying cellular senescence and its role in age-related diseases.
  • To investigate the use of conditioned medium from senescent hepatocytes to induce senescence in other hepatocytes.

Main Methods:

  • Replicative senescence was induced in L02 hepatocytes by prolonged cell culture (20 weeks).
  • Conditioned medium from these senescent hepatocytes was used to induce senescence in fresh hepatocyte cultures.
  • Senescence was validated using senescence-associated β-galactosidase (SA-β-gal) staining, EdU incorporation assays, and analysis of senescence-related proteins.

Main Results:

  • The conditioned medium from replicative senescent L02 hepatocytes successfully and rapidly induced senescence in recipient hepatocytes.
  • Validation assays confirmed the induction of senescence, demonstrating the efficacy of the developed model.
  • The study provides example results and discusses potential applications of this novel protocol.

Conclusions:

  • A rapid and effective in vitro model for inducing hepatocyte senescence using conditioned medium has been developed.
  • This model serves as a valuable tool for deconvoluting the mechanisms of cellular senescence in age-related disease progression.
  • Further applications of this protocol can accelerate research into aging and associated pathologies.