Mouse splenocyte enrichment strategies via negative selection for broadened single-cell transcriptomics

Area of Science:

• Immunology
• Cell Biology
• Genomics

Background:

• Mammalian splenic tissue contains numerous immune cells, predominantly lymphocytes.
• High lymphocyte abundance can obscure the detection of low-abundance immune cell populations in analyses.
• Accurate characterization of all immune subsets is crucial for understanding immune responses.

Purpose of the Study:

• To develop a protocol for enriching minority immune cell populations from mouse spleen.
• To enable a more comprehensive analysis of the splenic immune landscape.
• To improve the resolution of single-cell transcriptomic studies.

Main Methods:

• Utilizing immunomagnetic negative depletion to remove abundant cell types.
• Generating an untouched, enriched fraction of rare immune cells.
• Controlled repopulation of enriched cells with untouched splenocytes.
• Validation using flow cytometry and single-cell transcriptomic analysis.

Main Results:

• Successful enrichment of low-abundance immune cell populations.
• Generation of an untouched, viable enriched cell fraction.
• Demonstrated broadened cellular landscape in single-cell transcriptomic clustering analysis.
• Validation of cell integrity and population representation via flow cytometry.

Conclusions:

• The described protocol effectively enriches rare immune cells from mouse spleen.
• This method enhances the cellular landscape revealed by single-cell transcriptomics.
• The protocol provides a valuable tool for detailed immunological studies.