Precision Culture Scaling to Establish High-Throughput Vasculogenesis Models

Hydrogel-based 3D cell cultures can recapitulate (patho)physiological phenomena ex vivo. However, due to their complex multifactorial regulation, adapting these tissue and disease models for high-throughput screening workflows remains challenging. In this study, a new precision culture scaling (PCS-X) methodology combines statistical techniques (design of experiment and multiple linear regression) with automated, parallelized experiments and analyses to customize hydrogel-based vasculogenesis cultures using human umbilical vein endothelial cells and retinal microvascular endothelial cells. Variations of cell density, growth factor supplementation, and media composition are systematically explored to induce vasculogenesis in endothelial mono- and cocultures with mesenchymal stromal cells or retinal microvascular pericytes in 384-well plate formats. The developed cultures are shown to respond to vasculogenesis inhibitors in a compound- and dose-dependent manner, demonstrating the scope and power of PCS-X in creating parallelized tissue and disease models for drug discovery and individualized therapies. Adapting 3D cell cultures for high-throughput screening workflows remains challenging. Precision culture scaling (PCS-X) effectively tailors 3D hydrogel-based vasculogenesis models for drug response profiling, integrating automated processes and analyses with Design of Experiment (DOE) and Multiple Linear Regression (MLR). PCS-X paves the way for hydrogel-based high-throughput 3D tissue and disease models as needed to advance drug discovery and individualized therapies. image