An In Vitro Model of Macrophage Senescence.
Grasiela Torres1, Utkarsh Tripathi1, Ivan Salladay-Perez1
1Microbiology, Immunology, Molecular Genetics, University of California Los Angeles, Los Angeles, California.
Current Protocols
|April 22, 2026
Summary
This study details a reproducible method for generating senescent macrophages from murine bone marrow. The protocol includes cryopreservation and induction of senescence via genotoxic stress, useful for studying innate immunity in aging.
Area of Science:
- Immunology
- Cell Biology
- Aging Research
Background:
- Macrophages are crucial innate immune cells implicated in aging.
- Cellular senescence, a state of irreversible cell cycle arrest, contributes to age-related pathologies.
- Investigating macrophage senescence is vital for understanding aging mechanisms.
Purpose of the Study:
- To establish a reproducible in vitro protocol for generating senescent murine macrophages.
- To provide a scalable method for macrophage culture and senescence induction.
- To facilitate research into the role of macrophage senescence in aging and disease.
Main Methods:
- Harvesting and culturing murine bone marrow-derived macrophages.
- Cryopreservation of bone marrow cells at -80°C for long-term storage.
- Inducing macrophage senescence using genotoxic stress (ionizing radiation or doxorubicin).
Main Results:
- Senescent macrophages displayed irreversible cell cycle arrest and characteristic senescence markers.
- Upregulation of senescence-associated markers like Cdkn1a was observed.
- Secretion of senescence-associated secretory phenotype (SASP) factors was confirmed.
Conclusions:
- The developed protocol provides a robust model for studying macrophage senescence.
- This model aids in understanding the innate immune system's contribution to aging.
- It offers mechanistic insights into age-related diseases driven by senescent macrophages.
