Detection and Characterization of Senescent Cells with Flow Cytometry

Area of Science:

• Cell Biology
• Immunology
• Biochemistry

Background:

• Cellular senescence is a vital process in human physiology, implicated in development, healing, and age-related diseases.
• Existing methods for identifying senescent cells, like SA-β-Gal, have limitations including subjectivity and complexity.
• Flow cytometry (FC) offers a reliable single-cell analysis method, with recent advancements including lipofuscin detection using GLF16.

Purpose of the Study:

• To present detailed protocols for detecting senescent cells using GLF16 in flow cytometry.
• To validate these protocols in human myeloma cell lines (NCI-H929, L363) and clinical bone marrow samples.
• To expand the application of FC in senescence research and clinical diagnostics.

Main Methods:

• Induction of cellular senescence using hydrogen peroxide (H₂O₂).
• Staining of senescent cells with GLF16 for flow cytometry analysis.
• Development of a specific gating strategy for accurate senescent cell identification.
• Application of protocols to bone marrow samples from multiple myeloma patients after red blood cell lysis.

Main Results:

• Detailed protocols for GLF16-based senescent cell detection via FC are provided.
• Successful identification of senescent cells in human myeloma cell lines.
• Demonstrated applicability of the method to clinical bone marrow samples, including patients with multiple myeloma.

Conclusions:

• The developed GLF16/FC protocols offer a robust and accurate method for senescent cell detection.
• These techniques enhance the utility of flow cytometry in senescence research.
• The protocols may serve as valuable surrogate tests in clinical diagnostics for senescence-related conditions.