Department of Neuropathology, Regensburg University Hospital, 93053 Regensburg, Germany.
Division of Interventional Immunology, Leibniz Institute for Immunotherapy, 93053 Regensburg, Germany.
Cells. 2023 Jul 14;12(14):1856. doi: 10.3390/cells12141856.
Conventional 2D cultures are commonly used in cancer research though they come with limitations such as the lack of microenvironment or reduced cell heterogeneity. In this study, we investigated in what respect a scaffold-based (Matrigel™) 3D culture technique can ameliorate the limitations of 2D cultures. NGS-based bulk and single-cell sequencing of matched pairs of 2D and 3D models showed an altered transcription of key immune regulatory genes in around 36% of 3D models, indicating the reoccurrence of an immune suppressive phenotype. Changes included the presentation of different HLA surface molecules as well as cellular stressors. We also investigated the 3D tumor organoids in a co-culture setting with tumor-infiltrating lymphocytes (TILs). Of note, lymphocyte-mediated cell killing appeared less effective in clearing 3D models than their 2D counterparts. IFN-γ release, as well as live cell staining and proliferation analysis, pointed toward an elevated resistance of 3D models. In conclusion, we found that the scaffold-based (Matrigel™) 3D culture technique affects the transcriptional profile in a subset of GBM models. Thus, these models allow for depicting clinically relevant aspects of tumor-immune interaction, with the potential to explore immunotherapeutic approaches in an easily accessible in vitro system.
传统的二维培养常用于癌症研究，但存在局限性，例如缺乏微环境或细胞异质性降低。在这项研究中，我们研究了基于支架（Matrigel™）的 3D 培养技术在哪些方面可以改善二维培养的局限性。对配对的二维和三维模型进行基于 NGS 的批量和单细胞测序显示，大约 36%的三维模型中转录关键免疫调节基因的情况发生改变，表明免疫抑制表型再次出现。这些变化包括不同 HLA 表面分子的呈现以及细胞应激物。我们还在与肿瘤浸润淋巴细胞（TILs）共培养的情况下研究了 3D 肿瘤类器官。值得注意的是，与二维模型相比，淋巴细胞介导的细胞杀伤似乎对清除三维模型的效果较差。IFN-γ释放以及活细胞染色和增殖分析表明，三维模型的耐药性升高。总之，我们发现基于支架（Matrigel™）的 3D 培养技术会影响一部分 GBM 模型的转录谱。因此，这些模型能够描绘出肿瘤免疫相互作用的临床相关方面，并有可能在易于获取的体外系统中探索免疫治疗方法。
