Strocchi Silvia, Santandrea Giacomo, Zanetti Eleonora, Verna Giulio, Cusenza Vincenza Ylenia, Nicoli Davide, Fantini Valentina, Grieco Alessandra, Paci Massimiliano, Ciarrocchi Alessia, Sancisi Valentina
Translational Research Laboratory, Azienda Unità Sanitaria Locale-IRCCS di Reggio Emilia, 42122 Reggio Emilia, Italy.
Pathology Unit, Azienda Unità Sanitaria Locale-IRCCS di Reggio Emilia, 42122 Reggio Emilia, Italy.
Cancers (Basel). 2024 Dec 25;17(1):27. doi: 10.3390/cancers17010027.
BACKGROUND/OBJECTIVES: Despite the introduction of innovative therapeutics, lung cancer is still the leading cause of cancer-related death. For this reason, lung cancer still requires deep characterization to identify cellular and molecular targets that can be used to develop novel therapeutic strategies. Three-dimensional cellular models, including patient-derived organoids (PDOs), represent useful tools to study lung cancer biology and may be employed in the future as predictive tools in therapeutic decisions. However, the successful establishment of lung cancer organoids cultures that faithfully represent the respective patient tissues is still challenging due to low success rate and/or overgrowth of normal airway epithelial cells.
We set up a two-step protocol that allows for establishing both short-term and long-term 3D cultures, with different characteristics and success rates.
Cancer tissue-originated spheroids (CTOSs) show a 100% success rate and allow for the concomitant isolation of autologous tumor infiltrating leukocytes (TILs). On the contrary, PDOs can be expanded for a medium-long term and bio-banked but retain a lower success rate and the possibility of contamination with normal airway epithelial cells. To overcome these problems, we set up an optimal medium formulation and we implemented rigorous quality controls, leading to a substantial improvement in the success rate of tumoral PDO establishment.
Overall, this protocol guarantees flexibility and reliability, also providing useful guidelines for quality control checks to support different experimental settings. The setting up of a robust protocol for lung cancer PDO culture establishment and expansion is a key requirement for their employment both in cancer research and as predictive tools in clinical practice.
背景/目的：尽管引入了创新疗法，但肺癌仍是癌症相关死亡的主要原因。因此，肺癌仍需要深入表征，以确定可用于开发新型治疗策略的细胞和分子靶点。三维细胞模型，包括患者来源的类器官（PDO），是研究肺癌生物学的有用工具，未来可能会用作治疗决策中的预测工具。然而，由于成功率低和/或正常气道上皮细胞过度生长，成功建立忠实地代表各自患者组织的肺癌类器官培养物仍然具有挑战性。
我们建立了一个两步方案，可用于建立具有不同特征和成功率的短期和长期三维培养物。
癌组织来源的球体（CTOS）成功率为100%，并允许同时分离自体肿瘤浸润白细胞（TIL）。相反，PDO可以进行中长期扩增并保存于生物样本库中，但成功率较低，且存在被正常气道上皮细胞污染的可能性。为了克服这些问题，我们制定了最佳培养基配方并实施了严格的质量控制，从而使肿瘤性PDO建立的成功率有了显著提高。
总体而言，该方案保证了灵活性和可靠性，还为质量控制检查提供了有用的指导方针，以支持不同的实验设置。建立一个强大的肺癌PDO培养建立和扩增方案是其在癌症研究和临床实践中用作预测工具的关键要求。
