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

Gillian Dao Nyesiga*, Jeppe Lohfert Haslund-Vinding, Josephine Budde, Josefine Føns Lange, Nadja Blum, Kotryna Dukstaite, Lars Ohlsson, Tiit Mathiesen, Anders Woetmann, Frederik Vilhardt

*Corresponding author for this work

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Abstract

Background: Current challenges in meningioma treatment, including post-surgical complications and cognitive impairments, highlight the need for new treatment alternatives. Immunological interventions have shown promise. However, there is a knowledge gap in characterizing infiltrating immune cells in meningioma and their interplay. Further studies on immune cells in single-cell suspensions from digested meningioma tissues could identify targetable mechanisms for non-surgical treatment options with fewer side effects. This study aimed to optimize a protocol for faster digestion of meningioma tissues into viable single-cell suspensions and to identify infiltrating immune cell populations. Methods: We modified a commercial kit intended for whole skin dissociation to digest resected meningioma tissues into viable single-cell suspensions. Tumor-infiltrating immune cell populations were characterized using flow cytometry. Results: Flow cytometry analyses revealed that the digested tissue was composed of viable immune cells, including predominantly CD14+ macrophages and CD3+ T cells, with minor populations of CD56+ NK cells and CD19+ B cells. In both of the two patient samples tested, half of the tumor-associated macrophages were TIM-3+, with a small proportion co-expressing CD83. Women were more likely to have a lower proportion of immune cells, B cells, and NK cells. Female patients with a high proportion of immune cells had a higher proportion of macrophages. Conclusion: We successfully optimized a protocol for generating single-cell suspensions with viable immune cells from meningioma tissues, revealing infiltrating antigen-presenting cells with an immunosuppressive phenotype, and lymphocytes. This short protocol allows advanced analyses of tumor-infiltrating cells using techniques such as single-cell RNA sequencing and flow cytometry, which require live, dissociated cells.

Original languageEnglish
Article number3942
JournalCancers
Volume16
Issue number23
Number of pages19
ISSN2072-6694
DOIs
Publication statusPublished - 2024

Bibliographical note

Publisher Copyright:
© 2024 by the authors.

Keywords

  • brain tumor
  • immune cells
  • lymphocytes
  • macrophages
  • meningioma
  • single-cell suspension
  • T cells
  • TIM-3
  • tumor digestion
  • tumor-infiltrating cells

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