A Novel Microplate Format for Tumor Cell Cryopreservation and 3D Tumor Spheroids Formation to Streamline Pre-clinical Drug Testing

About InSphero's tumor cell cryopreservation platform

Drug discovery and safety assessment rely on predictive in vitro models, with a growing emphasis on multicellular tissues that reflect organotypic functionality. Traditional 2D tumor cell cultures, while common in early drug screening due to their ease and compatibility with high-throughput systems, fail to replicate the complex 3D structure and physiology of actual tumors. This limitation, along with the challenges of generating 3D tissues for each test, hampers research efforts.

To overcome these barriers, we have developed a novel platform featuring cryopreserved 3D tumor models in microplate format, suitable for high-content screening. This platform allows for rapid testing, with experiments starting within 3 to 5 days after thawing frozen cell suspensions and re-forming 3D spheroids. Cryopreservation enables large-batch production, ensuring consistent starting conditions for testing, enhancing data quality, and facilitating cross-group comparability.

The platform includes models of frequently used tumor cell lines, representing common solid tumors. Functionality tests using the apoptotic agent Staurosporine show varying spheroid morphologies—compact, less compact, and loose aggregates—reflecting tumor diversity. Staurosporine treatment produces dose-response profiles indicating growth inhibition, tumor shrinkage, or cell death, as measured by size profiling and ATP-based metabolic analysis. The assays have demonstrated robustness in plates stored for up to 6 months at -80°C.

In summary, this innovative approach addresses the limitations of 2D cultures by providing a high-throughput, in-plate frozen 3D cell culture platform. It combines physiologically relevant tumor models with a flexible assay system, promising to advance cancer drug discovery through more effective in vitro testing.

Download InSphero's poster about the tumor cell cryopreservation platform

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