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Published on: November 28, 2015
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.
Insights
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.
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.
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.
