PO.TB04.05 · 肿瘤生物学
解锁 3D：TumorMACS 培养基作为原代 2D 和 3D 肿瘤类器官培养的双用途解决方案
Unlocking 3D: TumorMACS media as a dual-purpose solution for primary 2D and 3D tumor organoid cultures
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：3D 细胞培养模型因能比传统 2D 培养更准确地模拟体内肿瘤环境，正在癌症研究中日益受到关注。这些模型提供了对肿瘤生物学的关键洞见，包括细胞间相互作用、耐药性和基因表达谱。通过提供更具生理相关性的环境，3D 培养增强了临床前研究的预测能力，促进了更有效癌症疗法的开发。因此，能够同时支持 2D 和 3D 生长的多功能、可重复且高效的培养基需求，在推进癌症研究和药物发现方面日益重要。传统培养基往往缺乏在 2D 与 3D 培养之间无缝过渡所需的多功能性，给工作流程效率带来挑战。
方法：我们最初使用各自的 TumorMACS 培养基在 2D 条件下培养原代结肠癌、肺癌和乳腺癌细胞。随后将这些细胞转移至 3D 培养，方法为生成 4x10^5 细胞/ml 的生长因子减少型 Matrigel 微滴（包埋培养），或使用含 2%(v/v) Matrigel 的 2x10^4 细胞/ml TumorMACS 培养基（悬浮培养）。对于每个细胞系，在其相应实体特异性 TumorMACS 培养基中补充 StemMACS Y-27632，并用于每周两次的换液，培养最长可达 14 天。将成熟的类肿瘤解离为单细胞，用于计算倍增时间，并使用 MACSQuant Analyzer 进行表面标志物分析。
结果：在所有评估的细胞培养中，从 2D 到 3D 培养的过渡均获成功。包埋培养的倍增时间约为结肠和乳腺细胞 3 天，肺细胞 4 天。尽管静态悬浮培养表现出相似的倍增时间，但变异性更大。包埋培养中生长的类肿瘤在尺寸上更均一，提示一致性更好，这对可靠的检测结果至关重要。然而，除了 2D 细胞扩增外，悬浮培养可作为在检测开始前更高效地扩大细胞数量的有吸引力的替代方案。
结论：TumorMACS 培养基的双用途能力实现了 2D 与 3D 培养之间的无缝过渡，提高了研究效率并加速了癌症疗法的开发。其使用简化了实验工作流程，并为专用类肿瘤培养基提供了一种经济高效的替代方案。
查看英文原文 English abstract
Background: 3D cell culture models are gaining more and more interest in cancer research by more accurately mimicking the in vivo tumor environment compared to traditional 2D cultures. These models provide critical insights into tumor biology, including cell-cell interactions, drug resistance, and gene expression profiles. By offering a more physiologically relevant context, 3D cultures enhance the predictive power of preclinical studies, facilitating the development of more effective cancer therapies. As such, the demand for versatile, reproducible, and efficient media that support both 2D and 3D growth is increasingly important in advancing cancer research and drug discovery. Traditional media often lack the versatility required for seamless transitions between 2D and 3D cultures, posing challenges in workflow efficiency.
Methods: We initially cultivated primary colon, lung and breast cancer cells in 2D using the respective TumorMACS media. These cells were transferred into 3D cultures by generating 4x10^5 cells/ml growth factor reduced Matrigel domes (embedded culture) or using 2x10^4 cells/ml TumorMACS medium with 2%(v/v) Matrigel (suspension culture). For each cell line the entity specific TumorMACS media was supplemented with StemMACS Y-27632 and used for bi-weekly media changes, for up to 14 days of culture. Mature tumoroids were dissociated into single cells for doubling time calculations and surface marker analysis using the MACSQuant Analyzer.
Results : The transition from 2D to 3D cultures was successful across all cell cultures evaluated. Embedded cultures showed doubling times of approximately 3 days for colon and breast cells, and 4 days for lung cells. Although static suspension cultures exhibited similar doubling times, they presented greater variability. The higher uniformity in size of tumoroids grown in embedded cultures suggest improved consistency, which is critical for reliable assay results. However, next to the expansion of cells in 2D, suspension cultures can be an attractive alternative to scale cell numbers more efficiently before the assay start.
Conclusion : TumorMACS media's dual-purpose capability enables seamless transitions between 2D and 3D cultures, enhancing research efficiency and accelerating the development of cancer therapies. Its use simplifies experimental workflows and offers a cost-effective alternative to specialized tumoroid media.
利益披露 Disclosure
L. Eich,
Miltenyi Biotec B.V. & Co. KG Employment.
M. Schneider,
Miltenyi Biotec B.V. & Co. KG Employment.
O. Hardt,
Miltenyi Biotec B.V. & Co. KG Employment.
D. Agorku,
Miltenyi Biotec B.V. & Co. KG Employment.
B. Theek,
Miltenyi Biotec B.V. & Co. KG Employment.