Verloy Ruben, Privat-Maldonado Angela, Van Audenaerde Jonas, Rovers Sophie, Zaryouh Hannah, De Waele Jorrit, Quatannens Delphine, Peeters Dieter, Roeyen Geert, Deben Christophe, Smits Evelien, Bogaerts Annemie
Research Group PLASMANT, Department of Chemistry, University of Antwerp, 2610 Antwerp, Belgium.
Center for Oncological Research (CORE), Integrated Personalized and Precision Oncology Network (IPPON), University of Antwerp, 2610 Antwerp, Belgium.
Cells. 2025 Mar 18;14(6):450. doi: 10.3390/cells14060450.
Pancreatic ductal adenocarcinoma (PDAC) presents significant treatment challenges due to its desmoplastic reaction, which impedes therapeutic effectiveness, highlighting the need for advanced vitro models to better mimic the complex tumor environment. The current three-dimensional co-culture models of fibroblasts and endothelial cells are lacking, which presents a challenge for performing more comprehensive in vitro research. Our study developed triple co-culture spheroid models using MiaPaCa-2 and BxPC-3 cancer cell lines, with RLT-PSC and hPSC21 pancreatic stellate cell lines and the endothelial cell line HMEC-1. These models were assessed through growth assays, multicolor flow cytometry to optimize cell ratios, cell viability assays to evaluate drug responses, and a tube formation assay with a spheroid-conditioned medium to examine angiogenesis. Our triple co-culture spheroids effectively replicate the PDAC microenvironment, showing significant variations in drug responses influenced by cellular composition, density, and spatial arrangement. The tube formation assay showcased the potential of our models to quantitatively assess a treatment-induced angiogenic response. These cost-effective triple-co-culture in vitro spheroid models provide vital insights into the PDAC microenvironment, significantly improving the quality of the in vitro evaluation of treatment responses.
胰腺导管腺癌（PDAC）因其促结缔组织增生反应而带来了重大的治疗挑战，这种反应会阻碍治疗效果，凸显了需要先进的体外模型来更好地模拟复杂的肿瘤环境。目前缺乏成纤维细胞和内皮细胞的三维共培养模型，这给进行更全面的体外研究带来了挑战。我们的研究使用MiaPaCa-2和BxPC-3癌细胞系、RLT-PSC和hPSC21胰腺星状细胞系以及内皮细胞系HMEC-1开发了三重共培养球体模型。通过生长测定、多色流式细胞术以优化细胞比例、细胞活力测定以评估药物反应以及用球体条件培养基进行的管形成测定来检查血管生成，对这些模型进行了评估。我们的三重共培养球体有效地复制了PDAC微环境，显示出受细胞组成、密度和空间排列影响的药物反应存在显著差异。管形成测定展示了我们的模型定量评估治疗诱导的血管生成反应的潜力。这些具有成本效益的三重共培养体外球体模型为PDAC微环境提供了重要见解，显著提高了治疗反应体外评估的质量。
