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Human Microglia Seize the Chance to be Different.

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    Summary

    Human microglia show distinct regional differences and phenotypes. This study characterized human microglia (huMG) using mass cytometry, revealing their unique profiles across brain regions for better understanding in health and disease.

    Area of Science:

    • Neuroscience
    • Immunology
    • Cell Biology

    Background:

    • Microglia are key innate immune cells in the central nervous system, vital for neural development and function.
    • While rodent microglia phenotypes are known, human microglia (huMG) heterogeneity remains largely uncharacterized due to procurement and preservation challenges.
    • Previous studies were limited in scale, hindering comprehensive analysis of huMG diversity.

    Purpose of the Study:

    • To comprehensively characterize human microglia heterogeneity and phenotypes across different brain regions.
    • To establish a reliable method for analyzing postmortem human microglia.
    • To differentiate human microglia from peripheral myeloid cells and compare them with fresh human microglia.

    Main Methods:

    • Multiplexed mass cytometry was employed to analyze 57 markers on 103-104 postmortem human microglia (huMG).

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  • huMG were isolated from up to 5 distinct brain regions across 9 donors.
  • A hybrid workflow combining Cytobank and R/Bioconductor facilitated multidimensional data analysis for detecting regional heterogeneity.

    • Main Results:

    • A distinct phenotypic signature for human microglia (huMG) was identified, differentiating them from peripheral myeloid cells.
    • The immunophenotype of postmortem huMG was found to be comparable to that of fresh human microglia.
    • Significant regional heterogeneity in microglia was detected across different brain areas.
    • High-dimensional, single-cell immunophenotyping enabled unambiguous profiling of huMG.

    Conclusions:

    • This study successfully characterized human microglia heterogeneity and regional phenotypes using advanced mass cytometry techniques.
    • The findings provide a foundation for understanding microglia function in both healthy and diseased states.
    • The developed methodologies enable large-scale, high-dimensional profiling of human microglia at the single-cell level.