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Flow Cytometry Analyses of Meningioma Immune Cell Composition Using a Short, Optimized Digestion Protocol.

Gillian Dao Nyesiga1,2, Jeppe Lohfert Haslund-Vinding2, Josephine Budde2

  • 1Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, 205 06 Malmo, Sweden.

Cancers
|December 17, 2024
PubMed
Summary

Researchers optimized a protocol to analyze immune cells in meningioma tumors. This method identifies immunosuppressive macrophages and T cells, paving the way for new non-surgical treatments.

Keywords:
T cellsTIM-3brain tumorimmune cellslymphocytesmacrophagesmeningiomasingle-cell suspensiontumor digestiontumor-infiltrating cells

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Area of Science:

  • Neuro-oncology
  • Immunology
  • Cell Biology

Background:

  • Meningioma treatment faces challenges like post-surgical complications and cognitive issues.
  • Immunological approaches offer potential but require understanding of tumor-infiltrating immune cells.
  • Characterizing these cells is crucial for developing targeted, non-surgical therapies with fewer side effects.

Purpose of the Study:

  • To optimize a rapid protocol for digesting meningioma tissues into viable single-cell suspensions.
  • To identify and characterize the infiltrating immune cell populations within meningioma.

Main Methods:

  • A modified commercial kit for skin dissociation was used to process resected meningioma tissues.
  • Flow cytometry was employed to analyze the composition of tumor-infiltrating immune cells.

Main Results:

  • The optimized protocol yielded viable single-cell suspensions rich in immune cells, primarily CD14+ macrophages and CD3+ T cells.
  • Minor populations of CD56+ NK cells and CD19+ B cells were also identified.
  • Tumor-associated macrophages frequently expressed TIM-3, indicating an immunosuppressive phenotype, with some co-expressing CD83.

Conclusions:

  • A protocol was successfully optimized for generating viable immune cell suspensions from meningioma.
  • The study identified infiltrating antigen-presenting cells with immunosuppressive properties and lymphocytes.
  • This rapid protocol enables advanced analyses like single-cell RNA sequencing and flow cytometry on live, dissociated cells.