Perier Magali, Wang Litan, Simonneau Marine, Ngo-Reymond Jacqueline, Guillermet-Guibert Julie, Lafond Maxime, Lafon Cyril
LabTAU, INSERM, Centre Léon Bérard, Université Claude Bernard Lyon 1, 69003 Lyon, France.
Centre de Recherches en Cancérologie de Toulouse (CRCT), CNRS-Inserm, Université de Toulouse, 2 av Hubert Curien, 31037 Toulouse, France.
Methods Protoc. 2025 Jul 7;8(4):75. doi: 10.3390/mps8040075.
Pancreatic adenocarcinoma remains one of the deadliest cancers, with limited treatment options and high chemoresistance. Traditional 2D cell cultures fail to accurately replicate the tumor architecture. Our study introduces three-dimensional (3D) pancreatic adenocarcinoma spheroid models using magnetic aggregation of pancreatic cancer cells and immortalized fibroblasts in either liquid culture medium or embedded in hydrogels. The spheroids' growth was characterized using optical imaging, while viability was assessed using ATP quantification and flow cytometry. Results demonstrated successful spheroid formation and growth. Further analysis suggested that on one hand, culture in liquid medium and ATP-based viability assessment are practical for initial experiments. On the other hand, hydrogel culture and flow cytometry, although being more resource- and labor-intensive, provided both a more reproducible and detailed viability analysis.
胰腺腺癌仍然是最致命的癌症之一,治疗选择有限且化疗耐药性高。传统的二维细胞培养无法准确复制肿瘤结构。我们的研究引入了三维(3D)胰腺腺癌球体模型,该模型通过在液体培养基中或嵌入水凝胶中的胰腺癌细胞和永生化成纤维细胞的磁性聚集来构建。使用光学成像对球体的生长进行表征,同时使用ATP定量和流式细胞术评估活力。结果表明成功形成并生长了球体。进一步分析表明,一方面,在液体培养基中培养和基于ATP的活力评估对于初始实验是实用的。另一方面,水凝胶培养和流式细胞术虽然资源和劳动力需求更大,但提供了更可重复和详细的活力分析。