肿瘤微环境复杂且动态，其特点是血管化不良、营养供应有限、缺氧和酸性pH。这种环境在推动癌症进展中起着至关重要的作用。然而，用于体外研究癌细胞生物学的标准细胞培养条件无法复制肿瘤的体内环境。最近，已经开发出与人类血浆非常相似的“生理”细胞培养基（例如 Plasmax、HPLM），并且更频繁地采用生理氧条件（1-8% O2）。尽管如此，可能需要进一步完善肿瘤特异性培养条件。在这项研究中，我们描述了基于小鼠胰腺导管腺癌（PDAC）肿瘤间质液的肿瘤微环境培养基（TMEM）的开发。使用 RNA 测序，我们发现，与类血浆条件（小鼠血浆培养基，pH 7.4， 5% 氧气）。具体来说，与细胞迁移、生物合成、血管生成和上皮间质转化相关的基因和途径的表达发生了改变，表明肿瘤样条件促进了转移表型和代谢重塑。使用功能测定来验证 RNA-seq 数据，我们证实，尽管细胞增殖减少，但 1.5%O2/TMEM 下的运动性有所增加。此外，通过测量葡萄糖摄取/乳酸产生和线粒体呼吸，发现了向糖酵解代谢的标志性转变。总而言之，这些发现表明，1.5%O2/TMEM 中的生长以与癌症生物学相关的方式改变了多种生物反应，并且更接近地模拟了培养中的标志性癌症表型。这凸显了在标准癌症研究中建立类似肿瘤微环境的条件的重要性。

The tumour microenvironment is complex and dynamic, characterized by poor vascularization, limited nutrient availability, hypoxia, and an acidic pH. This environment plays a critical role in driving cancer progression. However, standard cell culture conditions used to study cancer cell biology in vitro fail to replicate the in vivo environment of tumours. Recently, 'physiological' cell culture media that closely resemble human plasma have been developed (e.g., Plasmax, HPLM), along with more frequent adoption of physiological oxygen conditions (1-8% O2). Nonetheless, further refinement of tumour-specific culture conditions may be needed. In this study, we describe the development of a Tumour Microenvironment Medium (TMEM) based on murine pancreatic ductal adenocarcinoma (PDAC) tumour interstitial fluid. Using RNA-sequencing, we show that murine PDAC cells (KPCY) cultured in tumour-like conditions (TMEM, pH 7.0, 1.5% O2) exhibit profound differences in gene expression compared to plasma-like conditions (Mouse Plasma Medium, pH 7.4, 5% O2). Specifically, the expression of genes and pathways associated with cell migration, biosynthesis, angiogenesis, and epithelial-to-mesenchymal transition were altered, suggesting tumour-like conditions promote metastatic phenotypes and metabolic remodeling. Using functional assays to validate RNA-seq data, we confirmed increased motility at 1.5%O2/TMEM, despite reduced cell proliferation. Moreover, a hallmark shift to glycolytic metabolism was identified via measurement of glucose uptake/lactate production and mitochondrial respiration. Taken together, these findings demonstrate that growth in 1.5%O2/TMEM alters several biological responses in ways relevant to cancer biology, and more closely models hallmark cancerous phenotypes in culture. This highlights the importance of establishing tumour microenvironment-like conditions in standard cancer research.