Bolster your confidence with your 3D cellular aggregates assay with automated bulk spheroids and organoids isolation and sorting

Complex three-dimensional (3D) in vitro models, in particular spheroids, tumoroids and organoids, offer unprecedented means to yield highly predictable models of healthy and diseased tissues and organs. For instance, tumor spheroids and tumoroids that display hallmarks of solid tumor such as multicellular composition, spatial architecture, diffusion barrier, differential gene expression and drug resistance, are currently the model of choice in anti-tumor drug screenings. Hitherto, the preparation of such cellular aggregates for high throughput screening relies on the aggregation of a cell suspension at the bottom of each individual wells of round multi-well plates. Although this method is compatible with automated liquid handling, it raises concern about well-to-well sample homogeneity and cellular debris scattering (ca. dead cells that failed to aggregate often surround the spheroid at the bottom of each wells). 3D spheroid preparation for high throughput screen would greatly benefit from automated cellular aggregate isolation and manipulation from bulk culture. Here, we present an automated large-particle sorter and dispenser that enables rapid and standardize 3D spheroid sample preparation. By merging Scienion’s sciDROP NANO® low-volume non-contact dispensing technology and cellenONE® software, Cellenion has developed the spheroONE®, an innovative platform to isolate, inspect, sort and dispense large-particle such as 3D spheroids. Here, we highlight the capabilities of the platform by demonstrating our ability to isolate spheroids with high accuracy (up to 100% single spheroid per well) and sort them according to user-defined characteristics (size, morphology, and signal in up to 4 fluorescent channels). The preparation of assay-ready plates by isolating and dispensing spheroids prepared in bulk has the potential to significantly accelerate drug screening processes and improve their predictability. Interestingly, the dark field illumination implemented in the spheroONE shows distinct diffraction pattern for cysts and aggregates with a lumen, which present an unprecedented mean to select organoids/tumoroids. The system target holders are temperature controlled which enable to encapsulate cellular aggregates in multi-well plates pre-loaded with extracellular matrix hydrogel (i.e. Matrigel) ensuring it remains liquid during the isolation process and only solidifies once placed in an incubator. We strongly believe the spheroONE will become an enabling technology toward the establishment of complex in vitro 3D models to reduce/replace animal experimentation.