(1116-A) Unveiling Drug Responses in Liver Spheroids: Multiplexing 3D Cell-Based Assays and Imaging in a Microwell Platform
Monday, February 5, 2024
12:00 PM – 1:00 PM EST
Location: Exhibit Halls AB
Abstract: The critical need to minimize animal testing and reduce drug attrition rates has resulted in the development of more complex in vitro models. Notably, 3D cell cultures have emerged as a significant alternative due to their ability to mimic the complex in vivo microenvironment, providing more physiologically relevant results compared to traditional 2D cell cultures. Liver spheroids were among the initial applications of 3D models for toxicity assessment, significantly enhancing the safety and efficiency of potential drug candidates' evaluation.
However, the widespread adoption of 3D cell cultures faces challenges related to reproducibility, automation, and tissue loss using conventional technologies. To address them, we present an innovative solution - Gri3D®: a novel hydrogel microwell 96 wellplate, where each well features a microwell array in a cell-repellent hydrogel. This design promotes uniform cell seeding, efficient aggregation, and the formation of a single microtissue per microwell under suspension-like conditions. The resulting microtissues are positioned within the same focal plane, enabling simultaneous automated imaging and multiple on-plate assays. The platform also incorporates a unique pipetting port for safe media exchange. Importantly, the Gri3D® platform is fully automatable, allowing for liquid handling solutions to manage media exchanges, compound addition, and assays without manual intervention.
In this study, we grew Upcyte® hepatocyte spheroids simultaneously in Gri3D® and U-bottom 96 well plates to compare ease of use, performance in terms of volume, required steps and time, sensitivity, detection window, and scalability for multi-assays. After exposure, drug effects on viability and enzymatic activities were assessed using Promega assays (CellTiterGlo®-3D, CellToxTM Green and P450-GloTM) and Tecan’s Spark® Cyto multi-mode imaging plate reader, with readouts encompassing fluorescence and luminescence intensity measurements, and brightfield and fluorescence imaging in each well.
Our innovative 3D multiplexing approach allowed us to gain valuable insights into the impact of different drugs on CYP activity and microtissue viability, all within a single experiment. Adopting the Gri3D® platform and a compatible multimode plate reader, along with multiplexed cell-based assays, allowed for efficient simultaneous analysis of multiple factors, advancing our understanding of liver spheroid's response to drug exposure. This approach shows immense potential in addressing critical challenges related to compound assessment and drug screening on a larger scale using 3D cell cultures.