Pancreatic Cancer 3D Cell Line Organoids (CLOs) Maintain the Phenotypic Characteristics of Organoids and Accurately Reflect the Cellular Architecture and Heterogeneity In Vivo

Abstract

Pancreatic cancer is a highly lethal disease. Therapeutic resistance to chemotherapy is a major cause of treatment failure and recurrence in pancreatic cancer. Organoids derived from cancer stem cells (CSC) are promising models for the advancement of personalised therapeutic responses to inform clinical decisions. However, scaling-up of 3D organoids for high-throughput screening is time-consuming and costly. Here, we successfully developed organoid-derived cell lines (2.5D) from 3D organoids; the cells were then expanded and recapitulated back into organoids known as cell line organoids (CLOs). The 2.5D lines were cultured long term into 2D established cell lines for downstream comparison analysis. Experimental characterisation of the models revealed that the proliferation of CLOs was slightly faster than that of parental organoids. The therapeutic response to chemotherapeutic agents in 3D CLOs and organoids showed a similar responsive profile. Compared to 3D CLOs and organoids, 2D cell lines tended to be less responsive to all the drugs tested. Stem cell marker expression was higher in either 3D CLOs or organoids compared to 2D cell lines. An in vivo tumorigenicity study found CLOs form tumours at a similar rate to organoids and retain enhanced CSC marker expression, indicating the plasticity of CSCs within the in vivo microenvironment.

Mini-organs with big impact: Organoids in liver cancer studies

“…A 2D model was considered to be a vital model for drug screening and cancer research traditionally but it has limitations when it comes to recapitulating the heterogenicity of the tumor. Two-dimensional models have homogenous growth and proliferation and hence are not able to mimic the heterogeneous nature of the tumor [ 39 , 40 ]. On the other hand, organoids maintain their microenvironment and are better models for drug testing and determining drug response since tumor heterogeneity acts as a main challenge for any therapy [ 41 , 42 ].…”

“Organoids”: Insights from the First Issues

“…These cells could be expanded long-term and afterwards utilized again to grow organoids. The resulting cell line organoids (CLOs) showed a similar response to chemotherapeutic agents as the organoids they were derived of, making them a valuable tool in drug screening and personalized medicine [10]. In another paper, Li et al addressed the important issue of mimicking articular joint tissue in vitro [11].…”