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.
背景：当前脑膜瘤治疗面临的挑战，包括术后并发症和认知功能障碍，凸显了对新型替代疗法的需求。免疫干预已显示出潜力。然而，在脑膜瘤浸润免疫细胞的特征及其相互作用方面仍存在知识空白。对消化后脑膜瘤组织单细胞悬液中免疫细胞的进一步研究，可能为副作用更小的非手术治疗方案找到可靶向的机制。本研究旨在优化一种方案，以更快地将脑膜瘤组织消化为具有活性的单细胞悬液，并鉴定其中的浸润免疫细胞群。 方法：我们改良了一种用于全皮肤解离的商业试剂盒，以将切除的脑膜瘤组织消化为具有活性的单细胞悬液。使用流式细胞术对肿瘤浸润免疫细胞群进行表征。 结果：流式细胞术分析显示，消化后的组织由具有活性的免疫细胞组成，主要包括CD14+巨噬细胞和CD3+T细胞，以及少量的CD56+NK细胞和CD19+B细胞。在两个测试的患者样本中，半数肿瘤相关巨噬细胞为TIM-3+，其中一小部分共表达CD83。女性患者更可能具有较低比例的免疫细胞、B细胞和NK细胞。而免疫细胞比例高的女性患者，其巨噬细胞比例也更高。 结论：我们成功优化了一种从脑膜瘤组织中制备含有活性免疫细胞的单细胞悬液的方案，揭示了具有免疫抑制表型的浸润性抗原呈递细胞以及淋巴细胞。这一简短的方案使得能够利用单细胞RNA测序和流式细胞术等技术对肿瘤浸润细胞进行深入分析，这些技术均需要活的、解离的细胞。
肿瘤(癌症)患者之家