Scalable and Automated Three-Dimensional Cell Cultures for High-Throughput and High-Content Screening

Introduction:
More biologically relevant in vitro cell models with improved functionality are needed for high-throughput (HTS) and high-content screening (HCS). Three-dimensional (3D) culture mimicking extracellular matrix provides better in vivo -relevancy compared to 2D culture. Assay scalability and automation compatibility in 3D HTS/HCS is of essence, especially from cell model reproducibility and automated liquid handling compatibility. Animal-derived hydrogels often used in 3D assays have known challenges to fit HTS/HCS due to e.g., batch variation, solidification in room temperature and auto-fluorescence. Nanofibrillar cellulose (NFC) GrowDex® hydrogels are reproducible, animal-free, fully defined and automated liquid handling compatible. GrowDex hydrogels are shear-thinning and their stiffness can be tuned by adjusting the mixing ratio with cell media allowing fast and scalable 3D cell assay preparation for HTS and HCS.

Results:
GrowDex hydrogels have been shown to be a suitable extracellular matrix for culturing a wide range of 3D cell and tissue models. There are over 180 assay protocols for different cell types including primary, ES, iPS, patient-derived cells, biopsies and cell lines. Cells in GrowDex form spheroids or organoids and have been successfully screened against drug libraries. Several properties make GrowDex highly compatible with automated handling in HTS and HCS assays; it does not require cross-linking, it is non-autofluorescent, it is not temperature sensitive, it is well defined and being a shear-thinning material, it is easy and reproducible to dispense. The possibility to customize the matrix with wanted molecules provides full control to mimic the desired microenvironment for the cells. Finally, 3D cell cultures grown in GrowDex can be isolated and recovered for detailed downstream applications with the GrowDase™ cellulase enzyme treatment without impacting cell viability, phenotype or functionality.

Conclusions:
GrowDex hydrogels are animal free, easy-to-use hydrogel that mimics the extracellular matrix (ECM) and supports cell growth and differentiation with consistent results. GrowDex hydrogels are a proven solution for automated dispensing and HTS/HCS as a ready-to-use, reproducible and shear-thinning material.