3D培养越来越被应用于癌症研究中，因为它能够在模拟体内环境的条件下培养细胞。转移是癌症患者发病率和死亡率的主要原因，固体肿瘤转移主要发生在淋巴结中。目前，还没有能够在体外模拟前转移淋巴结微环境的技术。本研究中，我们制备了一种新型的细胞外基质——淋巴凝胶（Lymphogel），它源自淋巴结，模拟了转移性癌细胞的肿瘤微环境（TME）。我们通过各种功能性实验测试了新基质的适用性，并将结果与使用现有基质的结果进行了比较。我们使用了商业化和患者源性的原发性和转移性口腔舌鳞癌（OTSCC）细胞系。我们利用三种不同的基质（来自人子宫平滑肌瘤的Myogel，来自人前转移颈淋巴结的Lymphogel（h-LG），来自猪正常颈淋巴结的Lymphogel（p-LG））对这些细胞的增殖、粘附、迁移和侵袭进行了功能差异性研究。我们还进行了蛋白质组分析，以比较不同基质在功能特性上的差异。OTSCC细胞在不同基质上表现出不同的粘附和侵袭模式。转移细胞系在h-LG上的粘附能力有所提高，但基质对细胞侵袭的影响在细胞系之间波动无显著差异。蛋白质组分析显示，不同基质的蛋白质组成高度可变；Myogel含有618种蛋白质，p-LG含有1823种蛋白质，h-LG含有1520种蛋白质。三种基质的比较只发现了120种共有蛋白质。与每种基质的蛋白质组有关的细胞信号通路和过程的分析显示，Myogel与h-LG有相似性，但与p-LG的相似性较小。同样，与Myogel和h-LG相比，p-LG含有最少的与粘附相关的蛋白质。与粘附相关的独特蛋白质的数量最多的是h-LG。我们证明了人前转移颈淋巴结源基质适用于研究转移性OTSCC细胞。作为整体蛋白提取物，h-LG为体外癌细胞培养实验提供了新机会。 © 2023. BioMed Central Ltd., part of Springer Nature.

3D culture is increasingly used in cancer research, as it allows the growth of cells in an environment that mimics in vivo conditions. Metastases are the primary cause of morbidity and mortality in cancer patients, and solid tumour metastases are mostly located in lymph nodes. Currently, there are no techniques that model the pre-metastatic lymph node microenvironment in vitro. In this study, we prepared a novel extracellular matrix, Lymphogel, which is derived from lymph nodes, mimicking the tumour microenvironment (TME) of metastatic carcinoma cells. We tested the suitability of the new matrix in various functional experiments and compared the results with those obtained using existing matrices.We used both commercial and patient-derived primary and metastatic oral tongue squamous cell carcinoma (OTSCC) cell lines. We characterized the functional differences of these cells using three different matrices (human uterine leiomyoma-derived Myogel, human pre-metastatic neck lymph node-derived Lymphogel (h-LG), porcine normal neck lymph node-derived Lymphogel (p-LG) in proliferation, adhesion, migration and invasion assays. We also performed proteomic analyses to compare the different matrices in relation to their functional properties.OTSCC cells exhibited different adhesion and invasion patterns depending on the matrix. Metastatic cell lines showed improved ability to adhere to h-LG, but the effects of the matrices on cell invasion fluctuated non-significantly between the cell lines. Proteomic analyses showed that the protein composition between matrices was highly variable; Myogel contained 618, p-LG 1823 and h-LG 1520 different proteins. The comparison of all three matrices revealed only 120 common proteins. Analysis of cellular pathways and processes associated with proteomes of each matrix revealed similarities of Myogel with h-LG but less with p-LG. Similarly, p-LG contained the least adhesion-related proteins compared with Myogel and h-LG. The highest number of unique adhesion-related proteins was present in h-LG.We demonstrated that human pre-metastatic neck lymph node-derived matrix is suitable for studying metastatic OTSCC cells. As a whole-protein extract, h-LG provides new opportunities for in vitro carcinoma cell culture experiments.© 2023. BioMed Central Ltd., part of Springer Nature.