Selective Sorting of Senescent Cell Subpopulations Compatible with Downstream Genomics Applications

Area of Science:

• Cellular senescence research
• Genomics and epigenomics
• Biotechnology

Background:

• Cellular senescence is a complex state, making senescent cell identification difficult.
• Chromatin alterations are a hallmark of senescence, often compatible with mild cell fixation.
• Existing methods for senescent cell isolation are limited.

Purpose of the Study:

• To develop and detail a modified protocol for selective isolation of senescent cells using GLF16.
• To enable downstream genomics applications requiring specific fixation and permeabilization.
• To demonstrate the utility of the protocol for transcriptomics analysis.

Main Methods:

• Utilized the GLF16 compound for senescent cell identification.
• Developed two versions of a modified protocol for fluorescence-activated cell sorting (FACS) of senescent cells.
• Applied the sorted senescent cells to standard transcriptomics analysis.

Main Results:

• Successfully sorted a subpopulation of senescent fetal lung fibroblasts.
• Demonstrated the feasibility of using the sorted cells for transcriptomics.
• The protocol is adaptable for other genomics assays like CUT&RUN, CUT&Tag, ATAC-seq, and Hi-C/Micro-C.

Conclusions:

• The GLF16-based protocol provides an effective method for isolating senescent cells.
• This technique facilitates advanced genomics studies on senescent cell populations.
• The protocol's versatility supports a range of high-throughput sequencing applications.