(1229-B) Automated workflow for 3D cell model dispensing using spheroONE and phenotypic profiling using Pu·MA System EC

Abstract: Three-dimensional cell models have gained popularity, compared with traditional 2D models, because they better reproduce key aspects of human tissues and are amenable to a wide range of applications from basic research to pharmaceutical drug safety and efficacy testing. Methods for generating 3D models such as organoids, spheroids, and tumoroids have progressed to where scientists have the ability to recapitulate most human organs and cancer types ex vivo. However, the transition from 2D to 3D cell models has resulted in challenges related to sample handling and assay development and more sophisticated protocols and instrumentation are required. Here we present a novel automated workflow for phenotypic profiling of 3D models using spheroONE for automated sorting, isolation and dispensing of spheroids and Pu·MA system EC with low attachment flowchips for performing viability assays. The spheroONE is a large-particle sorter and dispenser that uses precision dispensing technology together with advanced image-based sorting capabilities to select and isolate single spheroids. The system was optimized to dispense size-selected spheroids into the protected sample chamber of Pu·MA System flowchips. The Pu·MA System EC with temperature, CO2 and relative humidity control was used to incubate the spheroids in the flowchips and perform viability assays using automated fluid transfers. The samples were stained in the flowchips with viability dyes and then imaged and analyzed. We used this workflow to analyze proliferation and viability of HCT116 colon cancer spheroids. Several thousand HCT116 spheroids were formed in a microwell cavity plate and collected into 5 ml of media. They were then aspirated by the spheroONE and 1 to 3 single 300 uM spheroids were dispensed into the protected sample chamber of flowchip samples wells. Spheroids were checked for viability before and after dispensing. The spheroids were treated for 24 to 72 hours with known toxic compounds and then stained with viability dyes. The spheroids were imaged in the flowchips and analyzed for number of Live and Dead cells. The spheroONE was able to dispense desired number of spheroids with a success rate of >84% and excellent spheroid health. Spheroid viability results were found to be consistent with known response of the compounds. This automated workflow using spheroONE and Pu·MA System EC for 3D cell model dispensing and phenotypic profiling eliminates sample disturbance, manual handling errors and provides consistent reproducible high-quality data. This platform is a valuable tool in a wide range of research areas including disease modeling, drug discovery and personalized medicine.