Abstract
In [1], the authors described a workflow for creating fit-for-purpose models in cell biology using a perfusion-based MPS interface for dynamicmicroenvironmental control. As part of this effort, non-small cell lung cancer lines LXFA 677 and LXFA 677-res1 (a Gefitinib-resistant derivative) [2] from CharlesRiver Laboratories (CRL, Freiburg, Germany) were separately cultured under static and perfused conditions for 4 days. Cells were treated with Gefitinib in aserial dilution ranging from 30 μM to 0.049 nM. These lines and the drug were chosen for their available in vitro and in vivo data, allowing performancecomparisons. Perfusion experiments utilized flow chambers from Vitroscope (Trondheim, Norway), with 2D cultures exposed to a constant cell culture mediumflow of 1.5 mL/min and 0.15 mL/min , maintaining stable conditions and a wall shear stress (WSS) of 2.3 mPa and 0.23 mPa, respectively.Viability was measured using CellTiter-Glo®. Here, viability data from static cultures treated with Gefitinib are compared to perfused cultures, using otherwisethe same protocol. These results are further evaluated against in vivo PDX models of the same cancer lines, showing that LXFA 677 is sensitive to treatmentwhile LXFA 677-res1 is resistant. The results show that the cell lines LXFA 677 and LXFA 677-res1 are highly sensitive to shear stress. Previous research hasshown similar results [3], yet the threshold at which the viability of (non-)small cell lung cancer cells is significantly affected is so far unknown. This dataset is an initial step toward improving the physiological relevance of in vitro cell cultures by exploring microenvironmental factors. Future efforts willextend this approach to additional cancer cell lines and drug classes while incorporating real-time monitoring of key environmental variables (oxygen and pH)to enhance model predictiveness. Additionally, single-cell sequencing from in vivo, static, and perfused cultures will be performed to assess microenvironmentalimpacts on cancer cells. Ultimately, the goal is to supplement and partially replace in vivo experiments with a higher-throughput in vitro alternative.