Targeting Conserved Pathways in 3D Spheroid Formation of Diverse Cell Types for Translational Application: Enhanced Functional and Antioxidant Capacity

Three-dimensional (3D) in vitro spheroid/organoid culture increasingly appears to better mimic physiological states than standard 2D systems. The biological consequence of 3D spheroids, however, differs for different cell types: pluripotent embryonic stem cells (ESCs), differentiation and loss stemness occur, while the converse is true somatic cancer cells. Despite such diverse consequences, there are likely conserved mechanisms governing spheroid formation across types that unknown but could be efficiently targeted translational application. To elucidate processes, we performed transcriptome analysis with functional validation on 2D- 3D-cultured mouse ESCs, mesenchymal stromal/stem (MSCs), At both transcriptomic levels, resulted commitment towards known cell-specific outcomes. Surprisingly all types, downregulation cholesterol synthesis pathway was found during formation, modulation concomitantly affecting consequences; similar results were seen human types. Furthermore, improved antioxidant capacity after further enhanced pathway. These findings demonstrate profound consequences value understanding formation.